Sumoylation of a meiosis‐specific RecA homolog, Lim15/Dmc1, via interaction with the small ubiquitin‐related modifier (SUMO)‐conjugating enzyme Ubc9

Abstract

Sumoylation is a post‐translational modification system that covalently attaches the small ubiquitin‐related modifier (SUMO) to target proteins. Ubc9 is required as the E2‐type enzyme for SUMO‐1 conjugation to targets. Here, we show that Ubc9 interacts with the meiosis‐specific RecA homolog, Lim15/Dmc1 in the basidiomycete Coprinus cinereus (CcLim15), and mediates sumoylation of CcLim15 during meiosis. In vitro protein–protein interaction assays revealed that CcUbc9 interacts with CcLim15 and binds to the C‐terminus (amino acids 105–347) of CcLim15, which includes the ATPase domain. Immunocytochemistry demonstrates that CcUbc9 and CcLim15 colocalize in the nuclei from the leptotene stage to the early pachytene stage during meiotic prophase I. Coimmunoprecipitation experiments indicate that CcUbc9 interacts with CcLim15 in vivo during meiotic prophase I. Furthermore, we show that CcLim15 is a target protein of sumoylation both in vivo and in vitro, and identify the C‐terminus (amino acids 105–347) of CcLim15 as the site of sumoylation in vitro. These results suggest that sumoylation is a candidate modulator of meiotic recombination via interaction between Ubc9 and Lim15/Dmc1.

SUMOylation is required for normal development of linear elements and wild-type meiotic recombination in Schizosaccharomyces pombe

“…In meiosis, various proteins involved in the processing of DSBs are known to be SUMOylated (Sacher et al 2006;Ho et al 2001;Watts et al 2007;Koshiyama et al 2006), and recently SUMOylation of proteins involved in meiotic pairing and synapsis was also reported (Cheng et al 2006;Hooker and Roeder 2006). In the budding yeast, the formation of the SC starts with the formation of axial elements consisting of Red1 and Hop1 which assemble along chromosomal axes containing a meiotic version of cohesin and topoisomerase II (Klein et al 1992(Klein et al , 1999Zickler and Kleckner 1999).…”

Coprinus cinereus Mer3 is required for synaptonemal complex formation during meiosis

“…Not only is its meiotic cell cycle long and naturally synchronous, but large populations of synchronous meiotic cells are also easy to obtain (Li et al 1999;Kües 2000;Kamada 2002;Casselton and Olesnicky 1998). These advantages have enabled us to isolate and characterize many synapsis-related proteins in C. cinereus (Nara et al 1999(Nara et al , 2001Hamada et al 2007;Ishizaki et al 2002;Namekawa et al 2003a, b, c;Yamaguchi et al 2004;Iwabata et al 2005;Koshiyama et al 2006;Sakamoto et al 2007;Sakaguchi et al 2007) and to demonstrate their roles in vivo.…”

Two X family DNA polymerases, λ and μ, in meiotic tissues of the basidiomycete, Coprinus cinereus

“…For some years, we have investigated meiosis-related proteins and their relationships to meiotic chromosome pairing and recombination using the meiocytes of the basidiomycete, C. cinereus (Koshiyama et al 2006;Iwabata et al 2005;Hamada et al 2002Hamada et al , 2007Namekawa et al 2005;Sakaguchi and Lu 1982;Lu et al 1988;Lu and Sakaguchi 1991;Matsuda et al 1993Matsuda et al , 1996Gomi and Sakaguchi 1994;Sawado and Sakaguchi 1997;Kitamura et al 1997;Nara et al 1999Nara et al , 2000Nara et al , 2001Ishizaki et al 2002;Namekawa et al 2003a-c;Yamaguchi et al 2004;Ishibashi et al 2006a-c). Like Lilium, the meiotic process is long and naturally synchronous.…”