BIOL0003 Lecture 04: Modifications of Mendel II

Gene Interaction

Last time – modification of Mendel;

1) Incomplete Dominance

2) Multiple Alleles

3) Lethal Alleles

4) Pleiotropy: one change, many effects

5) Sex limitation – different effects in males and females.

In fact sex limitation is an introduction to the idea that genes may interact to produce

a phenotype – eg sex limited pattern baldness; dominant, but shows effect only in

individual with a Y chromosome. Breast cancer – the mirror image of this.

Can see in experiments with mice which repeat Mendel’s dihybrid cross: and follow his

second law, of independent assortment, but sometimes with subtle modifications

because of gene interaction.

Reminder – Mendel dihybrid cross: simple – gives 9 3 3 1 ratio

Best examples come from coat colours in mice. Much worked on; both in classic

genetic terms and now as model of gene regulation in development. At least 5 major

gene loci, some with several alleles, interact to produce the range of coat colours found

in lab mice. Several alleles at each locus, in fact, but will only talk about two at each

here, for simplicity.

A locus. Wild-type mouse is "agouti" colour - a "salt-and-pepper" appearance. Each

hair; dark shaft with a yellow band.

A is wildtype and dominant. Non agouti phenotype a removes the yellow band, so that

aa mice lacking a genetic change at any other coat colour locus are black.

B locus. Codes for colour of hair. B codes for black pigment, b for brown. BB and Bb

mice are black; bb mice are brown. Consider a cross:

AA BB (agouti) x aa bb (brown).

F1 all Aa Bb (agouti)

Cross these together. Get the 9: 3: 3: 1 ratio expected for unlinked loci but because of

gene interaction get some unexpected phenotypes. Slide of cross. Slide of

cinnamon mouse.

In the following, a dash - stands for A or a, or B or b.

9 A- B- (agouti)

3 A- bb (cinnamon: brown hair with yellow fleck)

3 aa BB (black)

1 aa bb (brown)

The C locus. Codes for the presence or absence of pigment. CC or Cc animals have

pigment, cc have none - they are albino. The albino allele is epistatic to alleles at other

loci; ie it prevents them from expressing their effects. If an animal is cc, it makes no

difference what its genotype at the A or the B locus is; it will still be white, as no

pigment is made.
Imagine a cross:

BB CC (black) x bb cc (albino).

F1 All Bb Cc (black)

Cross these F1s together. Again, - stands for B or b; or C or c

9 B- C- (black)

3 bb C- (brown)

3 B- cc (albino) }

1 bb cc (albino) }

Ie a 9:3:4 ratio; the last two classes look the same as they have the epistatic albino

allele at the C locus that masks the phenotype at the B locus.

Various other loci – eg The D locus Controls the expression of the pigment produced

by the other coat colour loci. DD and Dd allow full expression of the other coat colours,

but dd is dilute: it makes the colours look pale and milky by producing an uneven

distribution of pigment in the hair shaft.

The S locus; controls pigment distribution on the body. SS or Ss are not spotted, but

ss is piebald - large patches of colour - say black and dilute black - on the same

individual. White spots often found, too. Must work early in development as the skin is

being produced.

Same genes also found in horses, cows, and some of in humans. 5 slides – horse,

tiger. Can make complex genotype statements – complex cat.

Might seem that looking at coat colour genes in mice is esoteric – but they can have

other effects, too: eg some are the same as those involved in human skin colour

variation, but others are involved in human diseases. Have a dominant spotted

phenotype in some children. The gene is identical to a receptor molecule on the cell

surface which when mutated becomes a cancer gene called kit. Also involved in

diabete, obesity and neurodegeneration, and skin cancer. Dozens of such genes

now known in mice.

Gene interaction easy to study in mice as can set up breeding expts. Harder in

humans, but some clear examples:

The Secretor phenotype, much used in forensic work. Those who are SeSe or Sese

produce ABO (strictly speaking ABH; as the "O" group does produce a substance "H")

blood group substances in their saliva and other bodily fluids such as semen. If an

individual has the secretor phenotype, and licks an envelope containing say a blackmail

threat then he will leave a clue as to his ABO blood-group. If he has not, he will not.

Ditto in rape cases. To do so, though, he must have particular alleles at two loci, the Se

locus and the ABO locus.

Gene interaction also importance in human disease. Classic case – sickle cell

anaemia. Turns out that mutations at other loci can both help and hinder it. Notable –

haemoglobin itself controlled by two groups of genes (will return to this). Adult

haemoglobin – two chains, an alpha and a beta, coiled round each other. Each one

coded for by its own locus. However, there are other haemoglobins active before birth –

necessary as they need to be more avid for oxygen than the adult form as the foetus
has to suck oxygen from its mother’s blood. Main one is Fetal Haemoglobin

(haemoglobin gamma plus hb alpha). In all babies this lasts for up to a few months

after birth, but drops rapidly.

However there are two mutations one found in Saudi Arabia and the other in Senegal

(home of ancestors of many African Americans) lead to Hereditary Persistence of

Foetal Haemoglobin HPFH. The presence of the extra haemoglobin into adulthood

much reduces the symptoms of sickle cell disease in SS homozygotes. Has an even

greater effect on thalassemia – another haemoglobin disease common in the Middle

East, which involves a deletion of a section of the molecule, rather than just the change

of one amino acid. Effect can be dramatic. Classic of gene interaction. Interesting –

only known treatment of sickle cell symptoms is to use the chemical hydroxyl-urea.

Originally not known why it works, but now know that it switches on the foetal

haemoglobin gene even in adults.

And now, CRISPR, trials under way to edit patient’s cells, and manipulate haemoglobin

genes to increase production of foetal haemoglobin; return to patient; has dramatic

effects in trials.

Have already seen that patients with the same SS mutation can have very different

symptoms; stroke, paralysis, heart disease – saw when discussing pleiotropy.

Complementation: a Special Case of Gene Interaction.

Sometimes we obtain a surprising result when crossing two individuals from different

families or stocks with what seems to be the same recessive phenotype. All their

offspring are wildtype!

Can produce characteristic ratios: Eg, foxglove normally has purple petals; but now

and again populations are found with a few individuals with white petals.

Within a population - simple Mendelian patterns suggesting that white is a recessive

just like, say, Mendel’s green peas.

Strange thing happens if you cross white plants from different populations? All the F1

plants are purple, not white; and there is a ratio of 9 purple to 7 white in the F2. What is

going on?

In fact – just as in the mouse coat colours - there are two loci involved; and

homozygosity for a white allele at either of them will produce a white plant. Ie there is a

chain of reactions producing purple, and a break at any point will stop the pigment

being produced. Use the usual convention that a dash (-) can represent either the

dominant or recessive allele in certain crosses where a single dominant allele is

present.

P1 white line 1 x white line 2

P1 w1w1 W2W2 x W1W1 w2w2

F1 All W1w1 W2w2 (purple) SELF-FERTILISE

F2 9 W1- W2- (purple) 9

3 W1- w2w2 (white) }

3 w1w1 W2- (white) }7

 1 w1w1 w2w2 (white) }

Ie evidence of complementation.

Complementation tests very useful in genetics; test whether two alleles at a locus are

involved, or two different loci.

Imagine that a blue-eyed fruit fly turns up in your cultures. Cross with wild-type (red);

they are recessive in simple Mendelian way. Cross them together to make a

homozygous line.

In a different line, to your surprise, a green-eyed fly – again a recessive - turns up.

Make that homozygous, too.

Are blue and green alleles at the same locus, or are different loci involved? Easy to

find out; cross the green homozygote with the blue homozygote. If they are alleles at

the same locus, the F1 will be blue or green, depending which one is dominant or it

could be blue-green if intermediate dominance).

But if two LOCI are involved, then the offspring would be heterozygous at both of

them and would have red eyes – the alleles would complement each other. If we

have many mutants – green, blue, pink, purple, puce, orange, etc can repeat again

and again and sort them into “complementation groups”.

This is how biochemical pathways were first worked out: led to realisation that

pathways actually exist – that may be several steps as different pathways cooperate

to make one substance.

Dros Wild-typeeye; red. Two new mutants then turned up – brown and scarlet. Each

was individually recessive to wildtype; but a complementation test showed that the

double heterozygote B+ S+ is wild-type red. Ie brown and scarlet are alleles at two

different loci. Now know that two separate pigments, coded for by different genetic

pathways, involved in dros eye colour; one makes brown pigment, one makes bright

scarlet, together they give red wildtype eye. Same result as the foxglove cross – and

the introduction to using genetics to disentangle biochemical pathways.

In other words, Mendel’s laws are correct, but have to be modified in various ways.

Can see just how complicated this gets when use a geneticist’s insight to look at

what might seem like a simply phenotype.

DEAFNESS: A MICROCOSM OF MODIFIED MENDELISM

Many different loci, some dominant, some recessive; many with pleotropic effects;

and good evidence of gene interaction. As Darwin himself noted – blue eyed white

cats are deaf!

Inheritance of deafness – first studied by Alexander Graham Bell; UCL student but

left because of illness; invented the telephone – but this was initially an apparatus to

help the deaf. His mother was deaf; and his father wrote a book on lip-reading. and

Bell married a deaf woman. Telephone – invented to help the deaf.

Hearing pathway has many steps; from outer ear, through middle with its bones, to
inner (the cochlea), and nerve transmission to brain, followed by interpretation of
sound. Hundreds of genes might be involved, more than 100 already found.

Recessive mutations at a single locus, GJB2 or Connexin 26, account for more than
half of all genetic cases in some, but not all populations. Despite the fact that more
than 20 loci have been described for non-syndromic autosomal recessive deafness
(DFNB), The gene involved in this type of deafness is encodes the gap junction
protein connexin 26(Cx26).

Connexins are transmembrane proteins that form channels allowing rapid transport
of ions or small molecules between cells. Connexin 26 mutation – leads to too much
potassium; hair cells in cochlea die; profound deafness, sometimes before child
learns to speak. Has more than a hundred different alleles – ie multiple alleles as in
cystic fibrosis. Nearly all recessive, but some dominant.

Remarkable – last few weeks; gene therapy for mice lacking hair cells has have
connexion mutation – hearing cilia grow back again!

Some lethals - Classic example – Samaritans of West Bank of Jordan, around town
of Nablus – once abundant (good Samaritan story), now reduced to around 600.
Recessive. Gene now known to make a MYOSIN molecule – again, involved in
muscle; presumably a failure of the hair cells in the inner ear, but for a different
reason. Homozygotes for the aberrant allele (just like brachydactyly)– die before
birth.

Pleiotropic effects. Deafness itself often a pleiotropic effect of a gene with many

other major effects: “syndromic deafness”; ie a pleiotropic effect. Even albinism –

can cause deafness, as inner ear has melanin which it needs for function; if absent,

then deaf (explains deaf blue-eyed white cat, also). – Also many of the spotting genes

in mice ass with Waardenburg syndrome – people who may have white blaze of hair,

sometimes have one brown and one blue eye – and may suffer from severe deafness.

The gene involved reads message off DNA; fails to produce melanin in some places –
melanin important in brain and hearing; failure to lay down causes deafness. Has a

mouse coat colour model,

Also, some genes whose major phenotype is deafness also have

unexpected pleiotropic effects themselves – eg Usher syndrome (failure of myosin)

leads to blindness as myosin involved in retina.

Complementation; deafness the classic case of gene interaction. Deaf people often

marry each other -–and may expect to have deaf children; but, most of the time, they

do not – as they are homozygous for recessives at different loci (as in the foxglove

example above). Deafness often genetic, showing recessive inheritance within a

particular family. Deaf people often marry - but in about four such marriages out of five,

all their children have normal hearing. Why?

There are many different gene loci affecting hearing - some in the outer ear, some in

the malleus incus and stapes, some in the nerve connections to the brain. A break in

the chain for any of them will lead to deafness.

Sound -> A -> B -> C -> D -> E -> F -> Perception of sound.

First deaf individual -> AA -> BB -> cc -> DD -> EE -> FF ->

Second: -> AA -> BB -> CC -> DD -> ee -> FF ->

Offspring: -> AA -> BB -> Cc -> DD -> Ee -> FF ->

Ie they are heterozygotes at both C and E, and can hear. Loci C and E show

complementation: results of cross show that deafness is not due simply to alleles

segregating at a single locus.

Of course – some deafnesses are environmentally caused; eg

shortage of thyroid hormone while developing – “cretinism” –

once widespread as in Derbyshire (“Derbyshire neck”),

Switzerland. Cured by adding iodine to salt. Some caused by

biological agents from outside – eg syphilis-infected mother, baby born deaf; rubella

(German measles) also. That disease becoming common as some refuse the MMR

vaccine.

But also a grey area in which gene and environment interact. Eg – well known that

intense noise can damage hearing; but, black people (much melanin in skin and

perhaps in inner ear) better able to retain hearing in face of noise; eg Black and

White american workers in noisy environment – blacks retain hearing better; ie

interaction of gene and environment. Will discuss this in a later lecture, when we

begin to study the inheritance of characters you measure rather than count:

quantitative characters such as height, or obesity.

 KEY WORDS

GENE INTERACTION

PEACOCK

BREAST CANCER

DIHYBRID CROSS 1

DIHYBRID CROSS 2

DH CROSS 3

DH CROSS 4

LAB MICE

AGOUTI MOUSE

AGOUTI HAIR

BLACK MOUSE (NON AGOUTI)

B LOCUS – BROWN MOUSE

AGOUTI CROSS BROWN GIVES CINNAMON

CINNAMON MOUSE

C LOCUS – ALBINO

ALBINO CROSS – 9 3 4

PIE BALD MOUSE

MOUSE GENES IN HORSES

ALBINO TIGER

WHITE SPOTTED BABY

MICE CLOSE TO HUMANS TREE

MICE GENES IN HUMANS

SECRETOR DISTRIBUTION
HERED PERSISTENCE OF FETAL HB

PURPLE AND WHITE FOXGLOVES

COMPLEMENTATION CROSS; 9 TO 7 RATIO

COMPLEMENTATION GROUPS IN DROSOPHILA

DARWIN’S DEAF CAT

ALEXANDER GRAHAM BELL

STRUCTURE OF EAR

A FEW DEAFNESS LOCI

SYNDROMIC AND NON-SYNDROMIC FREQUENCIES

ORGANS AFFECTED IN SYNDROMIC

WARDENBERG SYNDROME

USHER SYNDROME

COMPLEMENTATION DEAF PEDIGREE

EXPLANATION

DERBYSHIRE NECK

IODINE DEFICIENCY MAP

RUBELLA

CHILD WITH GERMAN MEASLES

EFFECTS OF GERMAN MEASLES

MMR POSTER

INCOMP DOM

MULTIPLE ALLELES

PLEIOTROPY

GENE INTERACTION

COMPLEMENTATION
FINALLY, GENE ENVIRONMENT INTERACTION