In the first step of retroviral integration, integrase cleaves the linear viral DNA within its long terminal repeat (LTR) immediately 3' to the CA dinucleotide step, resulting in a reactive 3' OH on one strand and a 5' two base overhang on the complementary strand. In order to investigate the structural properties of the 3' end processing site within the Moloney murine leukemia virus (MMLV) LTR d(TCTTTCATT), a host-guest crystallographic method was employed to determine the structures of four self-complementary 16 bp oligonucleotides including LTR sequences (underlined), d(TTTCATTGCAATGAAA), d(CTTTCATTAATGAAAG), d(TCTTTCATATGAAAGA) and d(CACAATGATCATTGTG), the guests, complexed with the N-terminal fragment of MMLV reverse transcriptase, the host. The structures of the LTR-containing oligonucleotides were compared to those of non-LTR oligonucleotides crystallized in the same lattice. Properties unique to the CA dinucleotide step within the LTR sequence, independent of its position from the end of the duplex, include a positive roll angle and negative slide value. This propensity for the CA dinucleotide step within the MMLV LTR sequence to adopt only positive roll angles is likely influenced by the more rigid, invariable 3' and 5' flanking TT dinucleotide steps and may be important for specific recognition and/or cleavage by the MMLV integrase.

译文

:在逆转录病毒整合的第一步中,整合酶立即将其长末端重复序列(LTR)内的线性病毒DNA切割到CA二核苷酸步骤的3'处,从而在一条链上产生反应性3'OH,并在其上形成5'两个碱基的突出端互补链。为了研究莫洛尼鼠白血病病毒(MMLV)LTR d(TCTTTCATT)中3'末端加工位点的结构特性,采用宿主客体晶体学方法确定了四个自我互补的16 bp寡核苷酸的结构,包括LTR序列(带下划线),d(TTTCATTGCAATGAAA),d(CTTTCATTAATGAAAG),d(TCTTTCATATGAAAGA)和d(CACAATGATCATTGTG),来宾与MMLV逆转录酶的N端片段(宿主)复合。将含LTR的寡核苷酸的结构与在相同晶格中结晶的非LTR的寡核苷酸的结构进行比较。 LTR序列中CA二核苷酸步骤所特有的属性(独立于双链体末端的位置)包括正侧倾角和负侧滑值。 MMLV LTR序列中CA二核苷酸步骤仅采用正侧倾角的倾向可能受到更刚性,不变的3'和5'侧接TT二核苷酸步骤的影响,并且对于MMLV的特异性识别和/或裂解可能很重要整合。

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