Nigel Odhiambo Ojuang

261117704
21st November 2022

Structural Maintenance of Chromosomes Protein 4 (SMC 4)

1. Part of the conserved group of proteins that are the SMCs (Structural Maintenance of
Chromosomes), SMC 4 is a very important protein in the cell cycle. It is found in the
condensin complex along with other proteins that make up the complex. Not many
specifics are known about how these SMCs condense proteins, however, a few
experiments have been done to give us some idea of what is taking place. As an
individual protein, SMC 4, along with the assistance of ATP, coils and bends which in
turn causes the DNA to condense into a more suitable form for mitosis. (Kim and Lopero)
However, the SMC is found in a complex with other proteins and therefore works
together with SMC 2 and other non-SMCs to condense DNA. The complex forms
clusters (potentially multiple) in the presence of topoisomerase I which makes the DNA
coil up and become more compact. The complex is also responsible for converting nicked
forms or DNA into positive knotted forms in the presence of topoisomerase II. (UniProt).
From this description, we can see that SMC 4 is linked to the cell cycle through its
association to the condensin complex which gets DNA ready for mitosis.

Cluster formation by condensin complex

 2. According to the cycle base website, the SMC 4 gene is mostly expressed in the G2/M to
M phase of the cell cycle. This is consistent with what we believe its function is which is
assisting in the condensing of DNA from its interphase form. It starts getting recruited in
the transition phase into mitosis, serves its function and then we see a steady decline in its
expression after the metaphase. However, we see a sudden increase in the amount of
SMC 4 in the cell when we get back to G1 even though the mitotic portion of the cell
cycle is complete. Upon research, we find that deposition of a histone H3 variant/CENP-
A into centromeric chromatin (centromeres) happens separately from DNA replication.
This causes the condensin II to be called upon (contains SMC 4), with the help of a factor
named HJURP, loads the CENP-A into the centromeres during G1. All this is possible
because condensin II is localized in the nucleus throughout the cell cycle. (Barnhart-
Daily et al).Outside the loading the CENP-A, M-Cdk is a mediator of condensin II as it
activates it (also mediates the removal of another SMC structure, cohesion). We see proof
of this as SMC 4 and M-Cdk are expressed at the same time except at G1 (M-Cdk is not
used for mediation next to centromeres).

Expression of SMC 4 in the cell cycle

Expression of M-Cdk in the cell cycle
 3.Since SMC 4 is used by both condensin I and II, it is localised at two different points in
the cell in certain instances. During the S phase since the nuclear envelope has not yet
been broken, the SMC 4 that is localized in the nucleus next to the kinetochore belongs to
condensin II since it is always localized in the nucleus. (SophieBachellier et al.) It is not
being expressed strongly at this point. However, in the G2/M phase, the expression of
SMC 4 starts to rise and condensin I is called upon by the cell as well from the cytoplasm
where it was from the S phase. Once the wall of the nucleus comes apart, all the SMC 4
in the cell is localised at the nucleus. This is in line with the function SMC 4 which is
condensation of proteins for mitosis and that takes place in the nucleus. After the
chromatin has been condensed and the mitosis portion of the cell cycle is about almost
done, we see that SMC 4 is started to be expressed at a lesser degree meaning it is no
longer needed by chromatin. At this point, condensin I moves back to the cytoplasm
while condensin II stays in the nucleus meaning SMC 4 is being expressed in two areas
of the cell once again. The ability of the cell to deposit CENP-A in the centromeric
chromatin is due to SMC 4 constantly being localized in the nucleus. (Barnhart-Daily et
al).

Representation of SMC 4 during mitosis (black denoting no presence of SMC 4). If we focus on
the blue, we see the concentration of SMC 4 drops in the cytoplasm as it rises in the nucleus and
vice versa.(Walther, N et al)
Condensin, I marked in green and condensin II marked in red showing that SMC 4 is expressed
in 2 parts of the cell.(Research gate)
4.SMC 4 experiences cell cycle regulated modification due to one of the three modifications that
happens upstream of the protein is by PLK1(Serine/Threonine protein kinase PLK1). PLK1 is a
multi-purpose protein that is important in the M phase of the cell cycle as it facilitates or at least
aids in actions such as the regulation of centrosome maturation and spindle assembly, the
removal of cohesins from chromosome arms, the inactivation of anaphase-promoting
complex/cyclosome (APC/C) inhibitors, the regulation of mitotic exit and cytokinesis etc. In the
context of SMC 4 and the condensin complexes, we can hypothesize that PLK1 aids in stopping
aberrant chromatid separation. The aberrant chromosome separation can be triggered by
phosphorylated mutants of condensin (phosphorylated subunits of condensin). (Kumar, S et al.)
Therefore, the PLK-1 interacting checkpoint Helicase (PICH), which is a substrate of PLK1 on
the kinetochore causes PLK to bind which keeps the chromatids stable during metaphase. (Dutta,
A et al.)

Modification proteins of SMC 4

5.From the blast that was carried out on SMC 4 comparing it to the yeast genome, we discover
that we only get about a 37% identity as our highest match but have more or less the same main
function in both organisms (DNA folding). However, the folding of chromosomes in yeast
happens at a lesser degree due to mitosis being endonuclear. (SophieBallachier et al.). They
share 5 out of 6 conserved protein domains which shows a similar ancestor or convergent
evolution (Homologs) . The one protein that is not conserved does lead to a divergence in some
of their functions. Yeast is lacking the PRK03918 which is used in DNA repair while Human
SMC 4 is lacking the PRK01156 which is used for provisional segregation (done by
microtubules in human cells). Therefore, we can conclude that SMC 4 in humans does not
participate in provisional segregation while SMC 4 in budding yeast does not participate in DNA
repair which may contribute to a lower percent identity.

Conserved domains in homo sapiens

Conserved proteins in budding yeast

Top hits from blast

Citation:

1.Uniprot website fallback message. UniProt. (n.d.). Retrieved November 21, 2022, from
https://www.uniprot.org/uniprotkb/Q9NTJ3/entry

2. Kim, H. J., & Loparo, J. J. (2016, January 4). Multistep Assembly of DNA condensation
clusters by SMC. Nature News. Retrieved November 21, 2022, from
https://www.nature.com/articles/ncomms10200

3. Barnhart-Dailey, M. C., Trivedi, P., Stukenberg, P. T., & Foltz, D. R. (2017, January
1). HJURP interaction with the condensin II complex during G1 promotes CENP-A
Deposition. Molecular biology of the cell. Retrieved November 21, 2022, from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5221629/

4., SophieBachellier-BassiaPersonEnvelope, a, Olivier Gadala1, 1, Gaëllen Bourouta, Ulf

Nehrbassb, (2008, January 11). Cell cycle-dependent kinetochore localization of
condensin complex in saccharomyces cerevisiae. Journal of Structural Biology. Retrieved
November 21, 2022, from
https://www.sciencedirect.com/science/article/pii/S1047847708000038#section0005

5. Dutta, A., Das, A., Bisht, D., Arya, V., & Muthuswami, R. (2022, October 18). PLK-1
interacting checkpoint helicase, Pich, mediates cellular oxidative stress response. MDPI.
Retrieved November 21, 2022, from https://www.mdpi.com/2075-4655/6/4/36/
6. Kumar, S., Sharma, G., Chakraborty, C., Sharma, A. R., & Kim, J. (2017, June 6). Regulatory
functional territory of PLK-1 and their substrates beyond mitosis. Oncotarget. Retrieved
November 21, 2022, from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5514964/

Pictures:

1. Kim, H. J., & Loparo, J. J. (2016, January 4). Multistep Assembly of DNA condensation
clusters by SMC. Nature News. Retrieved November 21, 2022, from
https://www.nature.com/articles/ncomms10200

2.Walther, N., Hossain, M. J., Politi, A. Z., Koch, B., Kueblbeck, M., Ødegård-Fougner, Ø.,
Lampe, M., & Ellenberg, J. (2018, July 2). A quantitative map of human condensins provides
new insights into mitotic chromosome architecture. Journal of Cell Biology. Retrieved November
21, 2022, from https://rupress.org/JCB/article/217/7/2309/39114/A-quantitative-map-of-human-
Condensins-provides

3.Localization of condensins I and II during the cell cycle. (a) hela ... (n.d.). Retrieved November
21, 2022, from https://www.researchgate.net/figure/Localization-of-condensins-I-and-II-during-
the-cell-cycle-A-HeLa-cells-were-fixed-with_fig4_8560213