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First published online 5 October 2004
doi: 10.1242/jcs.01417

Journal of Cell Science 117, 5427-5435 (2004)
Published by The Company of Biologists 2004
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Research Article

Caenorhabditis elegans UBC-2 functions with the anaphase-promoting complex but also has other activities

Tyralynn Frazier1,*, Diane Shakes2,*, Uma Hota1 and Lynn Boyd1,{ddagger}

1 Department of Biological Sciences, University of Alabama in Huntsville, Huntsville, AL 35899, USA
2 Department of Biology, College of William and Mary, PO Box 8795, Williamsburg, VA 23187, USA

{ddagger} Author for correspondence (e-mail: boydl{at}uah.edu)

Accepted 26 July 2004

The anaphase-promoting complex or cyclosome (APC/C) is a multi-subunit ubiquitin ligase that regulates the eukaryotic cell cycle. APC/C belongs to the RING finger class of ubiquitin ligases that function by interacting with a ubiquitin-conjugating enzyme (Ubc), thus inciting the Ubc to transfer ubiquitin onto a target protein. Extensive studies with APC/C in other organisms have identified several possible Ubcs that might function as partners for APC/C. This report presents phenotypic and biochemical evidence showing that, in Caenorhabditis elegans, UBC-2 interacts specifically with the APC/C. This conclusion is based on three lines of evidence: first, the RNAi phenotype of ubc-2 is indistinguishable from RNAi phenotypes of APC/C subunits; second, RNAi of ubc-2 but not other Ubcs enhances the phenotype of hypomorphic APC/C mutants; third, purified UBC-2 and APC-11, the RING finger subunit of the APC/C, show robust ubiquitination activity in in vitro assays. APC-11 interaction is specific for UBC-2 as ubiquitination is not seen when APC-11 is combined other C. elegans Ubcs. As expected from the Ubc that functions with the APC/C, ubc-2(RNAi) produces metaphase blocks in both mitotic germ cells and in meiotic divisions of post-fertilization oocytes. In addition, ubc-2(RNAi) results in two germline phenotypes that appear to be unrelated to the APC/C: an expanded transition zone indicative of a pre-pachytene meiotic arrest and endo-reduplicated oocytes indicative of a problem in ovulation or oocyte-soma interactions.

Key words: APC/C, E2 ubiquitin-conjugating enzyme, RING finger, E3 ubiquitin ligase, Ubiquitination, Metaphase-to-anaphase transition






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