Carteau S, Mouscadet J F, Goulaouic H, Subra F, Auclair C
Laboratoire de Physicochimie et Pharmacologie des Macromolécules Biologiques, CNRS URA 147, Institut Gustave Roussy, Villejuif, France.
Biochem Pharmacol. 1994 May 18;47(10):1821-6. doi: 10.1016/0006-2952(94)90311-5.
In search of potential inhibitors of integration of retroviral DNA into host cells genome, we have investigated the effect of the external DNA binder netropsin on the in vitro insertion of long terminal repeat (LTR) ends of Moloney murine leukemia virus (M.MuLV) as catalysed by integrase purified from baculovirus strain expression vector. In agreement with the preferential binding of netropsin to A+T rich sequences, footprinting experiments have shown that this drug selectively binds to the 5'-TTTCAT LTR end sequence which is included in the DNA binding site of integrase. This feature results in the potent inhibition of both reactions involved in the insertion process, namely, nucleolytic cleavage and strand transfer. The relation between netropsin binding to A+T rich region of M.MuLV LTR end and inhibition of insertion is strongly suggested from the inability of the drug to inhibit the insertion of HIV U3 LTR end which displays a G+C rich sequence. Selective inhibition of integration of viral DNA appears to be feasible using drugs recognizing LTR end sequences.
为寻找逆转录病毒DNA整合到宿主细胞基因组中的潜在抑制剂，我们研究了外源DNA结合剂纺锤菌素对莫洛尼鼠白血病病毒（M.MuLV）长末端重复序列（LTR）末端体外插入的影响，该插入反应由从杆状病毒株表达载体中纯化的整合酶催化。与纺锤菌素优先结合富含A+T序列一致，足迹实验表明该药物选择性地结合到整合酶DNA结合位点中包含的5'-TTTCAT LTR末端序列。这一特性导致插入过程中涉及的两个反应，即核酸裂解和链转移受到有效抑制。由于该药物无法抑制具有富含G+C序列的HIV U3 LTR末端的插入，因此强烈表明纺锤菌素与M.MuLV LTR末端富含A+T区域的结合与插入抑制之间存在关联。使用识别LTR末端序列的药物对病毒DNA整合进行选择性抑制似乎是可行的。
