uracil dna n-glycosylase promotes assembly of human centromere protein a尿嘧啶dna n-glycosylase促进人类着丝粒蛋白的组装.pdfVIP

Uracil DNA N-Glycosylase Promotes Assembly of Human Centromere Protein A 1 ¤ 2 3 4 5 Samantha G. Zeitlin * , Brian R. Chapados , Norman M. Baker , Caroline Tai , Geir Slupphaug , Jean Y. J. Wang4 1 Moores UCSD Cancer Center and Ludwig Institute for Cancer Research, University of California San Diego, La Jolla, California, United States of America, 2 Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, United States of America, 3 Department of Bioengineering, University of California San Diego, La Jolla, California, United States of America, 4 Moores UCSD Cancer Center, University of California San Diego, La Jolla, California, United States of America, 5 Faculty of Medicine, Norwegian University of Science and Technology (NTNU), Trondheim, Norway Abstract Uracil is removed from DNA by the conserved enzyme Uracil DNA N-glycosylase (UNG). Previously, we observed that inhibiting UNG in Xenopus egg extracts blocked assembly of CENP-A, a histone H3 variant. CENP-A is an essential protein in all species, since it is required for chromosome segregation during mitosis. Thus, the implication of UNG in CENP-A assembly implies that UNG would also be essential, but UNG mutants lacking catalytic activity are viable in all species. In this paper, we present evidence that UNG2 colocalizes with CENP-A and H2AX phosphorylation at centromeres in normally cycling cells. Reduction of UNG2 in human cells blocks CENP-A assembly, and results in reduced cell proliferation, associated with increased frequencies of mitotic abnormalities and rapid cell death. Overexpression of UNG2 induces high levels of CENP-A assembly in human cells. Using a multiphoton laser approach, we demonstrate that UNG2 is rapidly recruited to sites of