Article
- The EMBO Journal (2005) 24, 2172 - 2182
- doi:10.1038/sj.emboj.7600700
Published online: 2 June 2005
Subject Categories:
PIASy mediates SUMO-2 conjugation of Topoisomerase-II on mitotic chromosomes
Yoshiaki Azuma, Alexei Arnaoutov, Tadashi Anana and Mary Dasso
- Laboratory of Gene Regulation and Development, NICHD, NIH, Bethesda, MD, USA
Correspondence to:
Mary Dasso, Laboratory of Gene Regulation and Development, NICHD, NIH, Building 18, Room 106, 18 Library Drive, MSC 5431, Bethesda, MD 20892-5431, USA. Tel.: +1 301 402 1555; Fax: +1 301 402 1323; E-mail: mdasso@helix.nih.gov
aPresent address: Department of Pediatrics, Kumamoto University School of Medicine, Kumamoto, Japan
Received 4 November 2004; Accepted 9 May 2005
Abstract
Here we show that the PIASy protein is specifically required for mitotic modification of Topoisomerase-II by SUMO-2 conjugation in Xenopus egg extracts. PIASy was unique among the PIAS family members in its capacity to bind mitotic chromosomes and recruit Ubc9 onto chromatin. These properties were essential, since PIASy mutants that did not bind chromatin or failed to recruit Ubc9 were functionally inactive. We observed that PIASy depletion eliminated essentially all chromosomal accumulation of EGFP–SUMO-2-conjugated species, suggesting that it is the primary E3-like factor for mitotic chromosomal substrates of SUMO-2. PIASy-dependent SUMO-2-conjugated species were highly concentrated on the inner centromere, and inhibition of PIASy blocked anaphase sister chromatid segregation in egg extracts. Taken together, our observations suggest that PIASy is a critical regulator of mitotic SUMO-2 conjugation for Topoisomerase-II and other chromosomal substrates, and that its activity may have particular relevance for centromeric functions required for proper chromosome segregation.
Keywords:
- chromatin,
- mitosis,
- PIASy,
- SUMO,
- Topoisomerase-II
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