Pindyurin Alexey V, Boldyreva Lidiya V, Shloma Victor V, Kolesnikova Tatiana D, Pokholkova Galina V, Andreyeva Evgeniya N, Kozhevnikova Elena N, Ivanoschuk Igor G, Zarutskaya Ekaterina A, Demakov Sergey A, Gorchakov Andrey A, Belyaeva Elena S, Zhimulev Igor F
Institute of Cytology and Genetics of Siberian Division, Russian Academy of Sciences, Novosibirsk 630090, Russia.
J Cell Sci. 2008 May 15;121(Pt 10):1693-703. doi: 10.1242/jcs.018655. Epub 2008 Apr 29.
SUUR (Suppressor of Under-Replication) protein is responsible for late replication and, as a consequence, for DNA underreplication of intercalary and pericentric heterochromatin in Drosophila melanogaster polytene chromosomes. However, the mechanism by which SUUR slows down the replication process is not clear. To identify possible partners for SUUR we performed a yeast two-hybrid screen using full-length SUUR as bait. This identified HP1, the well-studied heterochromatin protein, as a strong SUUR interactor. Furthermore, we have determined that the central region of SUUR is necessary and sufficient for interaction with the C-terminal part of HP1, which contains the hinge and chromoshadow domains. In addition, recruitment of SUUR to ectopic HP1 sites on chromosomes provides evidence for their association in vivo. Indeed, we found that the distributions of SUUR and HP1 on polytene chromosomes are interdependent: both absence and overexpression of HP1 prevent SUUR from chromosomal binding, whereas SUUR overexpression causes redistribution of HP1 to numerous sites occupied by SUUR. Finally, HP1 binds to intercalary heterochromatin when histone methyltransferase activity of SU(VAR)3-9 is increased. We propose that interaction with HP1 is crucial for the association of SUUR with chromatin.
SUUR(复制不足抑制因子)蛋白负责后期复制,因此也负责黑腹果蝇多线染色体中间和着丝粒周围异染色质的DNA复制不足。然而,SUUR减缓复制过程的机制尚不清楚。为了确定SUUR可能的相互作用伙伴,我们以全长SUUR作为诱饵进行了酵母双杂交筛选。结果鉴定出HP1,一种研究充分的异染色质蛋白,是SUUR的强相互作用因子。此外,我们已经确定,SUUR的中央区域对于与HP1的C末端部分相互作用是必要且充分的,HP1的C末端部分包含铰链和染色体阴影结构域。此外,SUUR被招募到染色体上的异位HP1位点,为它们在体内的关联提供了证据。事实上,我们发现SUUR和HP1在多线染色体上的分布是相互依赖的:HP1的缺失和过表达都会阻止SUUR与染色体结合,而SUUR的过表达会导致HP1重新分布到SUUR占据的许多位点。最后,当SU(VAR)3-9的组蛋白甲基转移酶活性增加时,HP1会与中间异染色质结合。我们提出,与HP1的相互作用对于SUUR与染色质的关联至关重要。