Apoptosis — mechanism of apoptosis, intrinsic and extrinsic pathways Cell death + In humans, the rate of cell growth and cell death is balanced to maintain the weight of the body. wil0s its gas 18 10/30/2023Types of cell death Cell death can occur via several processes (about 11 types) : 1. Apoptosis 2. Necrosis 3, Autophagy 4. Entosis 5. Oncosis 6. Pyroptosis i. Two main mechanism of cell death + Apoptosis = “normal” or “programmed” cell death + Necrosis = “accidental” or “ordinary” cell death 10/30/2023Introduction of apoptosis >The word “apoptosis” comes from the ancient Greek, meaning the: “falling of petals from a flower” or “leaves fall off from a tree in autumn” >The term apoptosis (a-po-toe-sis) was first used in a now-classic paper by Kerr et al 1972 to describe a morphologically distinct form of cell death. Apoptosis definition * Apoptosis or programmed cell death (PCD) is a mode of cell death that occurs under normal physiological conditions and the cell is an active participant in its own demise (“cellular suicide’ * It is important for the development of multicellular organism (embryonic development) and homeostasis of their tissues (adult). 10/30/2023Importance of apoptosis ‘Apoptosis is a beneficial and important phenomenon: + Normal cell turn over + Tissue homeostasis + Induction and maintenance of immune tolerance + Development of the nervous system + Endocrine-dependent tissue atrophy (decrease in tissue mass of organ) + Elimination of activated, damaged and abnormal cells Apoptosis 1. Morphological features. ~ Volume reduction = Shrinking of cytoplasm = No loss of membrane integrity - Formation of apoptotic bodies = Condensation of chromatin & DNA fragmentation 2. Biochemical features Tightly regulated process Energy (ATP) - dependent (active process) Release of various factors into cytosol by mitochondria Activation of caspase cascade 10/30/2023Apoptosis... 3, Physiological significance Affects single cells or small clusters of cells Induced by physiological stimuli (lack of growth factors, DNA damage) Rapidly phagocytized by adjacent epithelial cells or macrophages No inflammatory response Apoptosis timing- Takes about 30 - 60min. Apoptosis: Intrinsic and extrinsic pathways INTRINSIC EXTRINSIC (Mitochondrial pathway) (Death receptor pathway) Involves release of cytochrome ¢ Activated by the engagement of (and other proteins) from ‘death receptors on cell surface ‘mitochondria eREHEEREREGEEDEEOET {Activation of caspases 10/30/2023Apoptosis: Intrinsic and extrinsic pathways. Mitochondrial pathway ; j FAS pathway oy oi, Bo dana Frecaspso S| DED-Death effector domain Caspases Caspases= Cysteinyl aspartate specific proteases + A family of intracellular cysteine proteases that playa pivotal role in the initiation and execution of apoptosis. + At least 14 different members of caspases in mammalian cells have been identified + All are synthesized as inactive proenzymes (zymogen) with 32-56 kDa 10/30/2023Caspase subgroups * To date, ten major caspases have been identified and broadly categorized into: 1. Signaling or | tor caspases (2, 8,9, 10) 2. Effector or Executioner caspases (3, 6, 7) 3. Inflammatory caspases (1, 4, 5) * The other caspases that have been identified include: Caspases 11, 12, 13, 14 + Central role in cascade of apoptotic events is played by caspase 3 Intrinsic Pathway Components: > Bcl-2 family proteins > Cytochrome ¢ > Adaptor proteins > Caspases 10/30/2023Intrinsic Pathway + The stimull that Initiate the Intrinsic pathway produce intracellular signals such as DNA damage, absence of certain growth factors, hormones and cytokines. + All of these stimuli cause changes in the mitochondrial outer membrane permeabilization (MO! * Release of pro-apoptotic proteins such as cytochrome ¢, Smac/DIABLO, Alf, endonuclease G and CAD (caspase activated DNAase) trom the inter-membrane space into the cytosol. * Cytochrome, binds and activates Apaf-2 as well as procaspase-9, forming an “apoptosome”. * Gaspase-9 activation, subsequent caspase-3 activation and cell jeath. Intrinsic Pathway... Be TOO Bion os Aaa ey dante {TRAZ- mitochondrial serine (PRE) a Ea NDDG ~Apoptotic DNAase 4 os zy SS 10/30/2023Intrinsic Pathway & Bcl2 family * Bcl-2 family members directly regulate the release of cytochrome -c. * This family contains both pro- and anti-apoptotic proteins. 1. Anti-apoptotic proteins include Bel-2, Bcl-x, Bcl-XL, Bcl-w 2. Pro-apoptotic proteins include Bax, Bak, Bid, Bad, Bim, Bik + The level between pro- and anti- apoptotic proteins determines if cytochrome cis released from the mitochondrion. Extrinsic Pathway Components: - Death Receptors - Death Ligands - Adaptor Proteins Caspases 10/30/2023Extrinsic Pathway + “Death receptors” that are members of the tumor necrosis factor (TNF) receptor superfamily. + Death receptors have a cytoplasmic domain of about 20 amino acids called the “death domain”. * This death domain plays a critical role in transmitting the death signal from the cell surface to th tracellular signaling pathways. Death Receptors & their ligands 6" The best characterized receptors & ligands corresponding death include: en FasR (CD95/APO1) Fast DR3 Apo3t DRA (TRAIL-R1) Apo2L DRS (TRAIL-R2) Apo2t TNFRL TNF-a ‘TNFR2 TNF-8 10/30/2023 10Extrinsic Pathway domain FAD} oo 4 2 hectare of spase 01400 Comrie) om : > Activation of caspase-8 > Activation of effector caspases > Apoptosis + apoptosis Apoptosis and cancer + Tumour cells can acquire resistance to apoptosis by 1. The expression of anti-apoptotic proteins or 2. by the down-regulation or mutation of pro-apoptotic proteins. + Resistance of tumour cells to apoptosis is an essential feature of cancer development. 10/30/2023 aApoptosis and cancer... Resistance mechanisms: + Expression of anti-apoptotic proteins (Bcl-2 over-expression in follicular 8-cell lymphoma; over-expression of IAPs in different types of cancers including neuroblastoma) * Inactivation of pro-apoptotic genes (BAX mutation; APAF-1in melanomas) * Alteration of p53 pathway (p53 mutation) * Altered survival signalling (alteration of PI3K/Akt pathway- for example PTEN deletion (phosphatase and tensin) 10/30/2023 12Regulation of cell cycle: Factors and genes regulating cell cycle * The cell cycle entails an ordered series of macromolecular events that lead to cell division + and the production of @©~—- two daughter cellseach ° ae J ©: == containing chromosomes identical to those of the parental cell. 10/30/2023The Cell Cycle Is an Ordered Series of Events Leading to Cell Replication + In cycling (replicating) somatic cells, cells synthesize RNAs and proteins during the G1 phase, + Preparing for DNA synthesis and chromosome replication during the S (synthesis) phase. + After progressing through the G2 phase, cells begin the complicated process of mitosis, also called the M (mitotic) phase, which is divided into several stages Regulated Protein Phosphorylation and Degradation Control Passage Through the Cell Cycle + The concentrations of the cyclins, the regulatory subunits of the heterodimeric protein kinases that control cell-cycle events, increase and decrease as cells progress through the cell cycle. * The catalytic subunits of these kinases, called cyclin-dependent kinases (CDKs), have no kinase activity unless they are associated with a cyclin. 10/30/202310/30/2023 Multiple CDks and Cyclins Regulate Passage of Mammalian Cells Through the Cell Cycle + Four CDKs are expressed at significant levels in most mammalian cells and play a role in regulating the cell cycle. + They are named as CDK1, 2, 4, and 6 + Cyclical changes in CDK activity are controlled mainly by cyclins + Cyclin Dand E are the mammalian | G1 phase cyclins. | + Cyclin A and cyclin B, which tuneion| in the S phase, G2, and early mitosis | G,-Cdk. cyclin D* Cdk4, Cdk6 G,/s-Cdk cyclin E Cdk2 s-Cdk cyclinA Cdk2, Cdk1** M-Cdk cyclin B Cdk1 * Gi-cyclinD — - help to pass the restriction point in late G1 + G1/S—cyclin E — Binds Cdks at the end of G1— commit DNA replication + S-cyclinA — - Binds Cdks during S phase - required for initiation of DNA replication + M-cyclinB — — promotes mitosis eventsCyclin-Cdk activation * Cyclin association + Kinase mediated phosphorylation rales S 1a y 8 Te Ff ee Regulation of the cyclin-Cdk complex 1. Post-translational modification 2. Inhibitors of CDKs 3. Cyclical proteolysis 4, Transcriptional regulation 10/30/2023| | t 1, Regulation of the cyclin-Cdk complex: post translational modification * Inhibition of Cdks by inhibitory phosphorylation * Inhibition: Phosphorylation by Weel kinase at TyrlS owe a * Cdc25 phosphatase dephosphorylates the p-TyriS and activates the Cdk 2. The inhibition of a cyclin-Cdk complex by Cdk inhibitor proteins (CKI) + CI (Cakinhibitor proteins): During Gi phase : The INK family, P15, p16, p18, end pis target to CDK4, & During all phases : The C1P/KIP family, p21, p27, snd p57 target to most CDKs + Binding of CXI renders Cek inactive &@& by conformation change and rearranged stricture of active ite 10/20/2023 uw3. Cyclical proteolysis regulates cyclin-Cdk activity a) SCF complex (Ubiquitin ligase) (Skp, Cullin, F-box containing complex) Cell cycle is regulated by SCF complex in G1 and S phase Targets:G1/S cyclins, some CKIs (such as p27Kip) ee oe F-box sa conserved domain that ls presentin numerous protein b) Regulated by APC complex in M phase APC: anaphase promoting complex Target: M cyclin APC complex (Ubiquitin ligase) contrat of protealyse by APCIC FRE sa @ el. 10/30/2023Regulation of G1, G1-S and S phases of cell cycle The G1 phase is a state of stable Cdk inactivity + To reacha stable G1 for growth, Cdk reactivation is prevented 1. Decline in M-cyclin and in Cdc20-APC 2. Decrease in M-cyclin transcription 3. Increase in Cdh1-APC* activity which targets to M- cyclins cetwen phase Cdh1 is a close relative of Cdc20 Cdh1-APC targets M-cyclins to inactivate M-Cdk after mitosis + Sict, a Cdk inhibitor (CK!), inactivates M-Cdk 10/30/2023Initiation of S phase in animal cells + In G1, accumulation of a CKI, and inhibition of cyclin gene ‘expression (M-cyclin) + In late G1, activation of G1-Cdk reverses the inhibitory state of G1 * G1-Cdk and G1/S-Cdk phosphorylate retinoblastoma (Rb), an inhibitor of cell cycle progression, and results in release of E2F Entry into mitosis inion wrote) coxsctvaing e€f62€° cant Inactive nt ry inact sctive Mca eee Snare ‘nce + M-cyclin accumulates by reduced degradation before M phase in embryonic cell cycles + Mz-cyclin increases at transcriptional level in most cell types 10/30/2023Inactivated M-cyclin-Cdk complex accumulates due to inhibitory phosphorylation Cdc25 phosphatase dephosphorylates and activates M-Cdk Polo kinase and active M-Cdk activate Cdc25 Active M-Cdk inactivates Weel Multiple roles of M-Cdk in mitosis Induce the assembly of mitotic spindle Ensure replicated chromosomes attach to the mitotic spindle Chromosome condensation Nuclear envelope breakdown Reorganization of the Golgi apparatus and endoplasmic reticulum 10/30/2023Exit from mitosis and start of G1 * Complex changes at the end of mitosis 1. The mitotic spindle must be dissassembled 2. Chromosomes decondensed 3. The nuclear envelope reformed Inactivation of M-Cdk is required for exit from mitosis Cde20-APC complex mediated ubiquitin- dependent proteolysis of M-cyclin DNA damage checkpoints and p53 activation * The DNA-damage checkpoint blocks progression through the cell cycle until the damage is repaired. + Damage to DNA can result from chemical agents and from irradiation with ultraviolet (UV) light or y-rays. * Arrest in G1 and S prevents copying of damaged bases, which would fix mutations in the genome. + Arrest in G2 allows DNA double-stranded breaks to be repaired before mitosis. 10/30/2023 10Tumor-suppressor protein, p53, contributes to arrest of cells with damaged DNA. Cells with functional p53 arrest in G1 and G2 when exposed to ~ irradiation Normally, p53 is targeted for ubiquitin-dependent degradation by Mdm2, a ubiquitin ligase The rapid degradation of p53 is inhibited by ATM and probably ATR, which phosphorylate “*” p53 ata site that interferes with binding by Mdm2. This increase the ability of p53 to activate transcription of specific genes that help the cell cope with DNA damage. One of these genes encodes p21 CIP, a generalized CIP that binds and inhibits all mammalian cyclin- CDK complexes. As a result, cells are arrested in G1 and G2 until the DNA damage is repaired and p53 and subsequently p21CIP levels fall 10/30/2023 aAn overview of the cell-cycle control system * Accelerators (cyclin-CDK) and brakes (checkpoints) oe =A | 10/30/2023 12Meiosis and genetic recombination + Meiosis increases the genetic variability in a population of organisms from one generation to the next. + Independent assortment allows maternal and paternal chromosomes to become shuffled during formation of the gametes + Genetic recombination (crossing-over) allows maternal and paternal alleles on a given chromosome to become shuffled as well. + By mixing maternal and paternal alleles between homologous chromosomes, meiosis generates organisms with novel genotypes and phenotypes. 10/30/2023Definition of recombination * Breaking and rejoining of two parental DNA molecules to produce new DNA molecules * Reciprocal recombination: new DNA molecules carry genetic information from both parental molecules. *Gene conversion: one way transfer of information, resulting in an allele on one parental chromosome being changed to the allele from the other homologous chromosome Gene conversion vs crossing over (recombination): (8) Gene conversion « pot a i a eee eee (€) Crossover — OT a > Ts vats ae F 10/30/2023+ Recombination is a remarkably precise process that normally occurs without the addition or loss of a single base pair. + Recombination depends on the complementary base sequences that exist between a single strand from one chromosome and the homologous strand of another chromosome. * The precision of recombination is further ensured by the involvement of DNA repair enzymes that fill gaps that develop during the exchange process. Biological Roles for Recombination 1. Generating new gene/allele combinations (crossing over during meiosis) 2. Generating new genes (e.g., Immuno-globulin rearrangement) 3. Integration of a specific DNA element (or virus) 4. DNA repair 10/30/2023Genetic recombination or Crossing Over * Crossing over may occur between non-sister chromatids at sites called chiasmata, + Crossing over: segments of non sister chromatids break and reattach to the other chromatid. * Chiasmata (chiasma) are where chromosomes touch each other and exchange genes (crossing over.) 10/30/2023Genetic recombination or Crossing Over Chiasmata - sites of crossing over, occur in synapsis. Exchange of genetic material between non- sister chromatids. Crossing over produces recombinant chromosomes. Types of Crossing Over Single Crossing Over: * In this type of crossing over only one chiasma is formed all along the length of a chromosome pair. Gametes- formed by this type of crossing over are called single cross over gametes. Double Crossing Over: + In this type two chiasmata are formed along the entire length of the chromosome leading to breakage and rejoin of chromatids at two points. The gametes produced are called double cross over gametes. 10/30/2023Multiple Crossing Over: + In this type more than two chiasmata are formed and thus crossing over occurs at more than two points on the same chromosome pair. The enzymes involved in crossing over 1) Recombinase is the major enzyme regulating recombination event (RADS1, DMC1). 2)Endonuclease 3)Ligase enzyme Endonuclease is responsible for breakage of 2 non-sister chromatids at corresponding sites. This is followed by the exchange of segments and finally the exchanged segments are joined or the gap is filled by ligase enzyme. 10/30/2023Proposed steps that occur during recombination in eukaryotic cells + Two DNA duplexes become aligned next to one another ‘Once they are aligned, an enzyme Spoii introduces a double-stranded break into one of the duplexes. + The gap is subsequently widened (resected) as indicated in step 2. + Resection may occur by the action of a5’ to 3” exonuclease. + The broken strands possess exposed single- stranded tails, each bearing a3’ OH terminus. + Meiotic recombination proceeds via binding of RADS1, and DMC1 to single- stranded DNA (ssDNA) substrates created ° after formation of programmed DNA double-strand breaks. a ‘+ RADiation sensitiveSi (RADS1) and DMC1 Disrupted Melotic cDNA1 (DMC1)recombinases, © mediate the search for homologous sequences + One of the single-stranded talls leaves its ‘own duplex and Invades the DNA molecule of ‘anonsister chromatid, hydrogen bonding with the complementary strand in the neighboring duplex (step 3). + The reciprocal exchange of DNA strands takes place 10/30/202310/30/2023 *+ The two duplexes are covalently linked to one another to form a joint molecule {or heteroduplex) that contains a pairof_ DNA crossovers, or Holliday Junctions, that flank the region of strand exchange (steps 4 and 5), These junctions are named after Robin ° Holliday, who proposed their existence in 1964, Linkage may move in one direction or another (an event known as branch Sa migration) by breaking the hydrogen i and reforming hydrogen bonds between (oS strands of the newly joined duplexes (step 5). e + To resolve the interconnected DNA molecules of the Holliday junctions, another round of DNA cleavage occur. * Depending on the particular DNA strands that are cleaved and ligated, two alternate products can be generated. 1, The two duplexes may contain only short stretches of genetic exchange, which represents a non crossover (step 6). 2. In the alternate pathway of breakage and ligation, the duplex of one DNA molecule is covalently joined to the duplex ofthe homologous molecule, creating a site of genetic recombination (.e., a crossover) (step 7). + Crossovers, which represent the — ——_— fusion of a maternal and paternal 4 ——. chromosome (step 7), during meiosis |, “F# YaoumeppictElectron microscope (EM) of a Holliday Junction with a few melted base pairs around junction FACTORS AFFECTING CROSS OVER Mutation: mutation reduces crossing over Temperature: high and low temperature variations increase the percentage of crossing over in certain parts of the chromosome. X-ray Effect: Xray irradiations increase crossing over near centromere. Agi older age increases the rate of crossing over. 10/30/2023Significance of Crossing-over 1. Produces new combinations of traits. 2. Through crossing over segments of homologous chromosomes are interchanged and hence provide origin of new characters and genetic variations. 3. Crossing over plays a very important role in the field of breeding to improve the varieties of plants and animals. Cross-over is more likely between distant genes than close genes. Crossing-over may occur at any point on chromosome arms Infrequent Frequent crossing-over crossing-over inthisregion inthis region 10/30/2023 10Linkage * It Is tendency of genes on a chromosome to remain together and passed as such in next generation. * It brings more parental types. + Strength of linkage between two genes increases if they are closely placed on a chromosome. + Ithelps to maintain a newly improved variety. + Linkage map and genetic maps are constructed on the basis of crossing over Linkage... + 2.genes on separate chromosomes assort independently at meiosis. * 2 genes far apart on the same chromosome can also assort independently at meiosis. + 2 genes close together on the same chromosome pair do not assort independently at meiosis. * Arecombination frequency << 50% between 2 genes shows. that they are linked. 10/30/2023Linkage... Unked genes don’t always stay linked. ‘These linkage groups can be separated by cros: prophase | of meiosis. over during, When crossing over occurs, the genes that were previously linked become unlinked, creating four different types of chromosomes (gametes). ‘The proportions are not equal'because crossing over does not occur in every cell during meiosis. 10/30/2023 12Meiosis Structure of Chromosomes * Diploid - A cell possessing two copies of each chromosome (human body cells). + Haploid - A cell possessing a single copy of each chromosome (human gametes). mologous chromosomes are made up of sister chromatids ied at the centromere. Homologous Homologous chromosomes aT a Sister Slates chromatide chromatids 10/30/202310/30/2023 How does meiosis compare to mitosis? + Only gametes undergo meiosis. All other cells divide by mitosis. + In meiosis, chromosomes are copied once, and the nucleus divides twice. + In mitosis, the chromosomes are copied once, and the nucleus divides once. How does meiosis compare to mitosis?.... + Meiosis produces haploid cells. Mitosis produces diploid cells.* Meiosis involves two distinct divisions, called Meiosis | and Melosis I * By the end of Meiosis II, the 1 diploid cell that entered meiosis has become 4 haploid cells Meiosis | reduces the number of chromosomes from diploid to haploid + Meiosis 1! produces four haploid daughter cells 10/30/2023Meiosis Phases * Meiosis involves the same four phases seen in mitosis 1. prophase 2. metaphase 3. anaphase 4. Telophase + They are repeated during both meiosis | and meiosis Il. + The period of time between meiosis | and meiosis II is called interkinesis. * No replication of DNA occurs during interkinesis because the DNA is already duplicated. Prophase | + Prophase | occupies more than 90% of the time required for meiosis + Chromosomes begin to condense + In synapsis, the 2 members of each homologous pair of chromosomes line up side-by-side, aligned gene by gene, to form a tetrad consisting of 4 chromatids + During synapsis, sometimes there is an exchange of homologous parts between ‘norrsister chromatids. This exchange is called crossing over + Each tetrad usually has one or more chiasmata, X-shaped regions where crossing over occurred 10/30/2023Metaphase! + At metaphase |, tetrads line up at the metaphase plate, with one chromosome facing each pole + Microtubules from one pole are attached to the kinetochore of one chromosome of each tetrad + Microtubules from the other pole are attached to the kinetochore of the other chromosome PROPHASE! METAPHASE! ANAPHASE! Homologous he (tedand oie pan Anaphase! + In anaphase |, pairs of homologous chromosomes separate + One chromosome moves toward each pole, guided by the spindle apparatus * Sister chromatids remain attached at the centromere and move as one unit toward the pole PROPHASE | METAPHASE) ANAPHASE | Sowa tonne, Homologous chomosomes Teadatne up Pe oftomaagoue ed and be para ‘hromoramen abit sichuoge seamen a= 4 10/30/202310/30/2023 Telophase | and Cytokinesis + In the beginning of telophase |, each half of the cell has a haploid set of chromosomes; each chromosome still consists of two sister chromatids * Cytokinesis usually occurs simultaneously, forming two haploid daughter cells + In animal cells, a cleavage furrow forms; in plant cells, a cell plate forms + No chromosome replication occurs between the end of meiosis | and the beginning of meiosis Il because the chromosomes are already replicated Prophase I + Meiosis I! is very similar to mitosis + In prophase Il, a spindle apparatus forms + Inlate prophase II, chromosomes (each still composed of two chromatids) move toward the metaphase plate Wigs SY AD.S.4FMetaphase I + At metaphase Il, the sister chromatids are at the metaphase plate + Because of crossing over in meiosis |, the two sister chromatids of each chromosome are no longer genetically identical + The kinetochores of sister chromatids attach to microtubules extending from opposite poles Anaphase I + At anaphase ll, the sister chromatids separate * The sister chromatids of each chromosome now move as two newly individual chromosomes toward opposite poles 10/30/2023Telophase Il and Cytokinesis + Intelophase Il, the chromosomes arrive at opposite poles ‘+ Nuelel form, and the chromosomes begin decondensing. + Cytokinests separates the cytoplasm + At the end of meiosis, there are four daughter cells, each with a haploid set of unreplicated chromosomes + Each daughter cell is genetically distinct from the others and from the parent cell A Comparison of Mitosis and Meiosis * Three events are unique to meiosis, and all three occur in meiosis | + Synapsis and crossing over in prophase I: Homologous chromosomes physically connect and exchange genetic information + At the metaphase plate, there are paired homologous chromosomes (tetrads), instead of individual replicated chromosomes + At anaphase | of meiosis, homologous pairs move toward ‘opposite poles of the cell. In anaphase 1! of meiosis, the sister chromatids separate 10/30/2023A Comparison Of Mitosis And Meiosis Nondisjunction Accidents during meiosis can alter chromosome number + Abnormal chromosome count is a result of nondisjunction Nonlauncion Weiss a “= + Either homologous \_ pairs fail to AR 2 Separate wot, GES) (GS during yy meiosis | JN comes (all) Call D 10/30/2023Fertilization after nondisjunction in the mother results in a zygote with an extra chromosome 1. Nondisjunction in autosomes 2. Nondisjunction in sex chromosomes 10/30/2023 101. Nondisjunction in autosomes Down Syndrome + Nondisjunction in Chromosome number 21 + Down syndrome is a genetic disease caused by an error during meiosis, + The chromatids in chromosome 21 do not separate, so one of the sex cells gets an extra copy of chromosome 21. + Down syndrome causes a number of health problems and learning difficulties, but many people with Down syndrome have fulfilling lives. Down Syndrome... *This karyotype shows three number 21 chromosomes *An extra copy of chromosome 21 causes Down syndrome Ce jean Kren gay Hw WY 10/30/2023 1The chance of having a Down syndrome child goes up with maternal age late wth Gown rome ‘penssowetay 2. Nondisjunction in sex chromosomes + Nondisjunction can also produce gametes with extra or missing sex chromosomes + Unusual numbers of sex chromosomes upset the genetic balance less than an unusual number of autosomes 10/30/2023 12ABNORMALITIES OF SEX CHROMOSOME NUMBER IN HUMANS. Sox Origin of Frequency Chromosomes Syndrome Nondisjunction In Population XxY Kinofelter Meiosis in ogg or Ee syndrome ‘sperm formation (ale) ' xy None Meiosis in sperm 3000 (normal male) formation ‘| Xx Metafemale Meiosis in eg or poanl sperm formation ; xo Tuner Meiosis in egg or fave) syndrome sperm formation (female) 10/30/2023 13Mitosis + Mitosis: -division of somatic (body) cells + Meiosis -division of gametes 10/30/2023Mitosis * Mitosis is the process by which new somatic cells are produced for: + Cell or organism growth + Replacing damaged or old cells. Thisis a complex process requiring different stages Mitosis. + Organisms grow by the addition of cells + In multicellular organism some of these cells perform functions different from other cells. + The process of a cell becoming different is differentiation. * Under normal conditions once an animal cell becomes specialized it can no longer form an entire organism + However plant cells are totipotent and any cell can form an entire plant. 10/30/2023Terminology * Chromatin - thin fibrous form of DNA and proteins, building block for a chromosome * Chromatid - one strand of the chromosome after it has. duplicated for cellular division * Sister chromatids - identical structures that result from chromosome replication, formed during S phase * Centromere - The part of a chromosome that links sister chromatids, site where kinetochore forms. + Kinetochore - A protein structure on chromatids where the spindle fibers attach during cell division to pull sister chromatids apart + Centriole & centrosome - A centriole is a barrel microtubule structure. Two centrioles arrange selves, perpendicularly to form a centrosome ae 7S Anatomy of a Chromosome * Centromere - point where sister chromatids are joined — together —tecatromere | p-short arm; upward * Qelong arm; \—learm] downward + Telomere-tips of chromosome telomere chromatids 10/30/2023Eukaryotic Chromosomes - linear chromosomes - every species has a different number of chromosomes ~ composed of chromatin — a complex of DNA and proteins > Heterochromatin refers to highly coiled regions where genes aren’t expressed. >Euchromatin refers to loosely coiled regions where genes can be expressed. Eukaryotic Chromosomes... + Chromosomes are very long and must be condensed to fit within the nucleus. + nucleosome — DNA wrapped around a core of 8 istone proteins * nucleosomes are spaced 200 nucleotides apart along the DNA + further coiling creates the 30-nm fiber or solenoid 10/30/2023Eukaryotic Chromosomes... + If an organism has the Diploid number (2n) it has two matching homologues per set. * One of the homologues comes from the mother (and has the mother’s DNA)... the other homologue comes from the father (and has the father’s DNA). + Most organisms are diploid. + Humans have 23 sets of chromosomes... therefore humans| have 46 total chromosomes... Advantages of having Diploid number of chromosomes * Protects against the loss of a single gene (and its protein) due ‘to mutation + Like having a “back-up” system * Pairs of like chromosomes are called homologous chromosomes + The human genome carries about 25,000 genes spread out over 24 (22+XX or XY) different types of chromosomes 10/30/2023Cell Cycle The ceil cycle is a sequence of cell growth and division. ion involves mitosis and cytokinesis. * Somatic cell di + Mitosis involves division of the chromosomes. * Cytokinesis involves division of the cytoplasm. + Mitosis without cytokinesis results in multinucleate cells. Cell Cycle... + The eukaryotic cell cycle is made up of a repeating pattern of growth, DNA replication, mitosis, and cytokinesis. + Typical animal cell cycle lasts about 24 hours. ‘Two main phases: Interphase (about 23 hrs, 30 min) Cell growth, productive activity, DNA replication M phase (about 30 min) Chromosomes segregated to daughter cells Cell divides into two 10/30/2023Cell Cycle... + Stages in eukaryotic cell cycle Interphase * First gap phase (G1) * Synthesis phase (S) * Second gap phase (G2) M phase + Mitosis + Cytokinesis Cell Cycle... 10/30/2023Interphase Constitutes about 90% of cell cycle length * G, (gap phase 1) — time of cell growth * S phase — synthesis of DNA (DNA replication) - 2 sister chromatids are produced * G, (gap phase 2) chromosomes condense Interphase... GlorGap1 +The cell just finished dividing so. in Gap 1 the cell is recovering from mitosis * Duplication of organelles + Numberof cytoplasmic components (macromolecules) doubled 10/30/2023Interphase... S or Synthesis stage * DNA synthesis occurs * DNA replication results in duplicated chromosomes \ccessory proteins are synthesised duplicated sister chromosome chromatids: (2 DNA double helices) Coyight © 2005 Pearcn Prentice Hale Interphase... G2orGap2 + Errors in DNA replication will be repaired Pia + Preparation for mitosis * Organelles are replicated. + Makes proteins necessary for cell division + More growth occurs. 10/30/2023MITosIS + Mitosis begins after G 2 and ends before G 1 + is the process in which a eukaryotic cell separates the chromosomes in its cell nucleus, into two identical sets in two daughter cells. a (LL ff, Prophase Duration (10 min) 1. Chromosomes condense -become visible under light microscope -2 sister chrom chromosomes joined at a centromere 2. Centrioles move to the opposite sides of the nucleus Nucleolus disappears Spindle fibres begins to form yy 10/30/2023 10Late Prophase or Prometaphase: Duration (10 min) + The nuclear membrane fragments and the microtubules invade the nuclear area + Kinetochores will get attached to the centromere + The spindle fibers become attached to the kinetochore. + Centrioles have moved to the opposite poles + The spindle is completely formed centrioles Microtubules form acomplet} spindle chromatids centrioles Metaphase Duration (10 min) * Chromosomes reach their highly condensed state + Centricies move to polar ends: and projects spindle fibers to ‘connect each chromosome + The spindle fibers begin to contract to the centromeres of the chromosomes, which are now arranged along the middle of the spindle. * The chromosomes are aligned at the metaphase plate 10/30/2023 1Anaphase Duration (3 min) In anaphase; +The paired chromosomes (sister chromatids) separate ‘+ Separated chromatids move to opposite pole + Partial division of cytoplasm begins + Cleavage furrow starts to form Telophase Duration (10 min). + Chromosomes are at the poles + Centrioles replicate * Nuclear membrane forms around each of the two sets of 46 chromosomes 10/30/2023 12Cytokinesis * Cytokinesis ~ cleavage of the cell into equal halves + inanimal cells — constriction of actin filaments produces a Colls return to cleavage furrow Oe ae * in plant cells — plasma membrane forms a cell plate between the nuclei + in fungi and some protists — mitosis occurs within the nucleus; division of the nucleus occurs with cytokirfesis Variations on the Theme Plant cells— + everything is similar except for cytokinesis, because plant cells have to break down and reform the cell wall. * Vesicles fuse near the metaphase plate to form a cell plate that grows outward to form a cell wall. Prokaryotes (no nucleus)— + binary fission * Single circular chromosome + DNA replicates and daughter “rings” unlink Simpler than mitosis 10/20/2024 13* Rate of DNA replication is same for all cells of a species + Same cycle length for same type of cells + Different cycle lengths for different types of cells e.g. cells in red bone marrow divide every second e.g. nerve cells stay in G1 indefinitely * Rate of cell division is under control (checkpoints, molecular brakes, etc.) CONTROL OF THE CELLCYCLE * Regulatory proteins called cyclins control the cell cycle at checkpoints: * G1 Checkpoint—decides whether or not cell will divide * S Checkpoint—determines if DNA has been properly replicated * Mitotic Spindle Checkpoint—ensures chromosomes are aligned at mitotic plate 10/30/2023 14