Impact of human immunodeficiency virus type 1 RNA dimerization on viral infectivity and of stem-loop B on RNA dimerization and reverse transcription and dissociation of dimerization from packaging
Impact of human immunodeficiency virus type 1 RNA dimerization on viral infectivity and of stem-loop B on RNA dimerization and reverse transcription and dissociation of dimerization from packaging
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DOI：
10.1128/jvi.74.12.5729-5735.2000
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发表时间：
2000-06-01
影响因子：
5.4
通讯作者：
Laughrea, M
中科院分区：
文献类型：
--
作者：
Shen, N;Jetté, L;Laughrea, M
吻环结构域（KLD）包括茎环，称为吻环或二聚化起始位点（DIS）发夹（人类免疫缺陷病毒1型毒株HIV-1（Lai）和HIV-1（Hxb 2）中的核苷酸[nt] 248至270），位于称为茎环B的12-nt茎内环的顶部（nt 243至247和271至277），破坏茎环B使基因组二聚化减少了约50%，使前病毒DNA合成减少了约85%，并且使二聚体基因组RNA的解离温度保持不变。逆转录的最受影响的步骤是正链DNA转移，减少了大约80%。与茎环B或DIS发夹的破坏相比，删除nt 241至256或200至256不会显著降低基因组二聚化。我们得出结论，KLD是非模块化的：茎环B和DIS发夹中的突变对基因组二聚化、逆转录和腺苷酸化具有相似的影响，并且也是“非加性的”;即，跨越这两种结构的较大缺失对基因组二聚化和双聚化具有相同的影响，就好像茎-环B强烈影响DIS发夹功能，反之亦然。接吻环回文中的C258 G颠换使基因组二聚化降低了约50%，病毒感染性降低了约1.4 log。两个突变，CGCG 261--> UUAA 261（产生较弱的回文）和Delta 241-256抑制突变各自能够减少基因组二聚化，但不影响基因组包装。
The kissing-loop domain (KLD) encompasses a stem-loop, named kissing-loop or dimerization initiation site (DIS) hairpin (nucleotides [nt] 248 to 270 in the human immunodeficiency virus type 1 strains HIV-1(Lai) and HIV-1(Hxb2)), seated on top of a 12-nt stem-internal loop called stem-loop B (nt 243 to 247 and 271 to 277), Destroying stem-loop B reduced genome dimerization by similar to 50% and proviral DNA synthesis by similar to 85% and left unchanged the dissociation temperature of dimeric genomic RNA, The most affected step of reverse transcription was plus-strand DNA transfer, which was reduced by similar to 80%. Deleting nt 241 to 256 or 200 to 256 did not reduce genome dimerization significantly more than the destruction of stem-loop B or the DIS hairpin. We conclude that the KLD is nonmodular: mutations in stem-loop B and in the DIS hairpin have similar effects on genome dimerization, reverse transcription, and encapsidation and are also "nonadditive"; i.e., a larger deletion spanning both of these structures has the same effects on genome dimerization and encapsidation as if stem-loop B strongly impacted DIS hairpin function and vice versa, A C258G transversion in the palindrome of the kissing-loop reduced genome dimerization by similar to 50% and viral infectivity by similar to 1.4 log, Two mutations, CGCG261-->UUAA261 (creating a weaker palindrome) and a Delta 241-256 suppressor mutation, were each able to reduce genome dimerization but leave genome packaging unaffected.