Zakharova O D, Martiyanov I V, Timofeeva O A, Kolocheva T I, Fournier M, Andreola M L, Maksakova G A, Yamkovoy V I, Litvak S, Tarrago-Litvak L, Nevinsky G A
Novosibirsk Institute of Bioorganic Chemistry, Russian Academy of Sciences, Novosibirsk.
Biochemistry. 1998 Sep 22;37(38):13343-8. doi: 10.1021/bi980239g.
The topography and functional implications of the complex formed in vitro between human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and its primer tRNALys3 were studied in this work. On the basis of previous results showing the high affinity both of the native primer, tRNALys3, as well as that of mismatched short oligonucleotide primers for HIV-1 RT, we synthesized chimeric primers containing tRNALys3 linked to U and T residues of different lengths. We found that the affinity of the oligonucleotide primers for HIV-1 RT is dramatically increased when linked to primer tRNA. Our results also show that in the tRNA.RT complex, before annealing tRNALys3 to the retroviral RNA genome, the 3'-terminal nucleotide of tRNALys3 is positioned at a distance of one nucleotide unit away from the template in the active polymerization site of the enzyme.
在本研究中，对1型人类免疫缺陷病毒（HIV-1）逆转录酶（RT）与其引物tRNALys3在体外形成的复合物的拓扑结构和功能意义进行了研究。基于先前的结果表明天然引物tRNALys3以及错配的短寡核苷酸引物对HIV-1 RT均具有高亲和力，我们合成了包含与不同长度的U和T残基相连的tRNALys3的嵌合引物。我们发现，当与引物tRNA相连时，寡核苷酸引物对HIV-1 RT的亲和力显著增加。我们的结果还表明，在tRNA.RT复合物中，在tRNALys3与逆转录病毒RNA基因组退火之前，tRNALys3的3'-末端核苷酸在酶的活性聚合位点中与模板相距一个核苷酸单位。
