a comparison of two single-stranded dna binding models by mutational analysis of apobec3g比较两个单链dna结合模型apobec3g的突变分析.pdfVIP

Biology 2012, 1, 260-276; doi:10.3390/biology1020260 OPEN ACCESS biology ISSN 2079-7737 /journal/biology Article A Comparison of Two Single-Stranded DNA Binding Models by Mutational Analysis of APOBEC3G Keisuke Shindo †, Ming Li, Phillip J. Gross, William L. Brown, Elena Harjes, Yongjian Lu, Hiroshi Matsuo * and Reuben S. Harris * Department of Biochemistry, Molecular Biology and Biophysics, Institute for Molecular Virology, University of Minnesota, Minneapolis, MN 55455, USA; E-Mails: shind009@kuhp.kyoto-u.ac.jp (K.S.); mingli@ (M.L.); gros0287@ (P.J.G.); brown344@ (W.L.B.); harje002@ (E.H.); luxxx079@ (Y.L.) † Present address: Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto 606-8507, Japan. * Authors to whom correspondence should be addressed; E-Mails: matsu029@ (H.M.); rsh@ (R.S.H.); Tel.: +1-612-625-6100. Received: 23 May 2012; in revised form: 1 July 2012 / Accepted: 14 July 2012 / Published: 2 August 2012 Abstract: APOBEC3G is the best known of several DNA cytosine deaminases that function to inhibit the replication of parasitic genetic elements including the lentivirus HIV. Several high-resolution structures of the APOBEC3G catalytic domain have been generated, but none reveal how this enzyme binds to substrate single-stranded DNA. Here, we constructed a panel of APOBEC3G amino acid substitution mutants and performed a series of biochemical, genetic, and structural assays to distinguish between “Brim” and