During cell division, the full set of chromosomes needs to separate accurately at anaphase to maintain complete genomic information in each daughter cell. Failure of proper chromosome segregation at anaphase leads to biased genomic information in daughter cells, which causes developmental defect and contributes to tumor progression. Precise structural organization of mitotic chromosomes is a key element in maintaining integrity of chromosome segregation, which is regulated by multiple posttranslational protein modification systems. Both genetic and biochemical studies indicate that the posttranslational protein modification by Small Ubiquitin-like Modifier (SUMO) modification pathway has an important role in normal progression of mitosis. I have found that mitotic specific SUMO-2 modification has a crucial role for completion of faithful chromosome segregation in anaphase in Xenopus egg extract assay system. This mitotic SUMO-2 modification specifically requires the activity of the PIASy protein that is a member of conserved PIAS family of E3. PIASy mediates SUMO-2 modification on multiple mitotic chromosomal proteins. Inhibition of SUMOylation in mitosis by either addition of mutant form of Ubc9, SUMO E2, or anti-PIASy neutralizing antibodies causes compromised chromosome segregation at anaphase. The major PIASy-mediated SUMO-2 substrate during mitosis is DMA topoisomerase II (Topoll), which is known to have an essential function in organizing mitotic chromosomes. Inhibition of SUMOylation alters the chromosomal association status of Topoll. I hypothesize that the PIASy mediated mitotic SUMOylation has an essential function in regulating the organization of mitotic chromosomes via its substrates, which plays a crucial role in chromosome segregation at anaphase. I propose to investigate a regulatory mechanism of PIASy specific SUMOylation pathway (Aim1) and a consequence of mitotic SUMOylation with determining the function of Topoll SUMOylation (Aim2) and identifying novel PIASy substrates (Aim3) by using Xenopus egg extracts as a model system. This investigation will demonstrate the function of SUMO-2 modification in organization of mitotic chromosomes in vertebrates. Determination of regulatory mechanisms of PIASy mediated SUMOylation will provide an invaluable information for understanding SUMOylation pathway that has an impact on diverse cellular physiological functions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM080278-04
Application #
7847549
Study Section
Special Emphasis Panel (ZRG1-NDT-K (01))
Program Officer
Hamlet, Michelle R
Project Start
2007-07-01
Project End
2012-06-30
Budget Start
2010-07-01
Budget End
2011-06-30
Support Year
4
Fiscal Year
2010
Total Cost
$368,101
Indirect Cost
Name
University of Kansas Lawrence
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
076248616
City
Lawrence
State
KS
Country
United States
Zip Code
66045
Kaur, Kawaljit; Park, Hyewon; Pandey, Nootan et al. (2017) Identification of a new small ubiquitin-like modifier (SUMO)-interacting motif in the E3 ligase PIASy. J Biol Chem 292:10230-10238
Yoshida, Makoto M; Ting, Lily; Gygi, Steven P et al. (2016) SUMOylation of DNA topoisomerase II? regulates histone H3 kinase Haspin and H3 phosphorylation in mitosis. J Cell Biol 213:665-78
Sridharan, Vinidhra; Park, Hyewon; Ryu, Hyunju et al. (2015) SUMOylation regulates polo-like kinase 1-interacting checkpoint helicase (PICH) during mitosis. J Biol Chem 290:3269-76
Ryu, Hyunju; Yoshida, Makoto M; Sridharan, Vinidhra et al. (2015) SUMOylation of the C-terminal domain of DNA topoisomerase II? regulates the centromeric localization of Claspin. Cell Cycle 14:2777-84
Ryu, Hyunju; Gygi, Steven P; Azuma, Yoshiaki et al. (2014) SUMOylation of Psmd1 controls Adrm1 interaction with the proteasome. Cell Rep 7:1842-8
Al-Ani, Gada; Briggs, Koan; Malik, Shuja Shafi et al. (2014) Quantitative determination of binding of ISWI to nucleosomes and DNA shows allosteric regulation of DNA binding by nucleotides. Biochemistry 53:4334-45
Sudharsan, Raghavi; Azuma, Yoshiaki (2012) The SUMO ligase PIAS1 regulates UV-induced apoptosis by recruiting Daxx to SUMOylated foci. J Cell Sci 125:5819-29
Ryu, Hyunju; Al-Ani, Gada; Deckert, Katelyn et al. (2010) PIASy mediates SUMO-2/3 conjugation of poly(ADP-ribose) polymerase 1 (PARP1) on mitotic chromosomes. J Biol Chem 285:14415-23
Ryu, Hyunju; Azuma, Yoshiaki (2010) Rod/Zw10 complex is required for PIASy-dependent centromeric SUMOylation. J Biol Chem 285:32576-85
Ryu, Hyunju; Furuta, Maiko; Kirkpatrick, Donald et al. (2010) PIASy-dependent SUMOylation regulates DNA topoisomerase IIalpha activity. J Cell Biol 191:783-94

Showing the most recent 10 out of 11 publications