| The cohesin complex is required to keep sister chromatids together until their separation during mitosis or meiosis II. I identified three kleisin subunits of the cohesin complex in the basidiomycete Coprinus cinereus, Rec8, Rad21.1, and Rad21.2. Paralog-specific antibodies were generated against each protein and used to characterize localization patterns by immunofluorescence microscopy during prophase I of meiosis. Results show that Rec8, Rad21.1, and Rad21.2 localize to meiotic chromatin during prophase I. Association of Rec8 on meiotic chromatin peaked between leptotene and diffuse diplotene, primarily localized to the chromatin core, while association of Rad21.1 peaked at diffuse diplotene and was primarily on chromatin loops. Using a mutant strain of spo11, I showed that association of Rec8 is Spo11-independent. In addition, I characterized localization of Rec8 and Rad21.1 in the absence of a sister chromatid and showed that both cohesin subunits localize to meiotic chromatin in the absence of a sister chromatid, suggesting roles for Rec8 and Rad21 independent of sister-chromatid cohesion. |
