Distinct Roles for Nucleic Acid in In Vitro Assembly of Purified Mason-Pfizer Monkey Virus CANC Proteins
Distinct Roles for Nucleic Acid in In Vitro Assembly of Purified Mason-Pfizer Monkey Virus CANC Proteins
复制标题
核酸在纯化梅森-辉瑞猴病毒 CANC 蛋白体外组装中的独特作用
作者：
P. Ulbrich;S. Haubova;M. Nermut;E. Hunter;M. Rumlová;T. Ruml
与其他逆转录病毒不同，梅森-辉瑞猴病毒（M-PMV）在细胞质中组装未成熟的衣壳。我们比较了最小的组装能力域从M-PMV和人类免疫缺陷病毒1型（HIV-1）在体外组装成病毒样颗粒的存在和不存在的核酸的能力。由Gag的衣壳和核衣壳结构域组成的融合蛋白（CANC）及其N端修饰的突变体（ΔProCANC）分别用于模拟病毒核心和未成熟颗粒的组装。与HIV-1相反，CANC有效地组装成圆柱形结构，M-PMV的相同结构域是组装不合格的。RNA或寡核苷酸的添加并不能弥补这一缺陷。相比之下，M-PMV ΔProCANC分子能够组装成球形颗粒，而HIV-1分子则形成球形和圆柱形。对于M-PMV，添加纯化的RNA增加了ΔProCANC形成球形颗粒的效率，无论是在总量还是完整球体的数量方面。测定掺入的RNA量，并且对于rRNA和MS 2-RNA，掺入的量与基因组RNA的量相似。寡核苷酸也刺激组装;然而，它们以痕量掺入ΔProCANC球形颗粒中，不能作为组装的化学计量结构组分。因此，寡核苷酸可以通过瞬时相互作用诱导有利于组装的构象变化，而较长的RNA似乎有利于球形颗粒的完全组装。
ABSTRACT In contrast to other retroviruses, Mason-Pfizer monkey virus (M-PMV) assembles immature capsids in the cytoplasm. We have compared the ability of minimal assembly-competent domains from M-PMV and human immunodeficiency virus type 1 (HIV-1) to assemble in vitro into virus-like particles in the presence and absence of nucleic acids. A fusion protein comprised of the capsid and nucleocapsid domains of Gag (CANC) and its N-terminally modified mutant (ΔProCANC) were used to mimic the assembly of the viral core and immature particles, respectively. In contrast to HIV-1, where CANC assembled efficiently into cylindrical structures, the same domains of M-PMV were assembly incompetent. The addition of RNA or oligonucleotides did not complement this defect. In contrast, the M-PMV ΔProCANC molecule was able to assemble into spherical particles, while that of HIV-1 formed both spheres and cylinders. For M-PMV, the addition of purified RNA increased the efficiency with which ΔProCANC formed spherical particles both in terms of the overall amount and the numbers of completed spheres. The amount of RNA incorporated was determined, and for both rRNA and MS2-RNA, quantities similar to that of genomic RNA were encapsidated. Oligonucleotides also stimulated assembly; however, they were incorporated into ΔProCANC spherical particles in trace amounts that could not serve as a stoichiometric structural component for assembly. Thus, oligonucleotides may, through a transient interaction, induce conformational changes that facilitate assembly, while longer RNAs appear to facilitate the complete assembly of spherical particles.
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DOI：
10.1073/pnas.85.22.8420
发表时间：
1988-11-01
影响因子：
11.1
作者：
GORELICK, RJ;HENDERSON, LE;REIN, A
通讯作者：
REIN, A
逆转录病毒 RNA 包装：综述。
DOI：
10.1007/978-3-7091-9326-6_49
发表时间：
1994
期刊：
Archives of virology. Supplementum
影响因子：
--
作者：
Rein,A
通讯作者：
Rein,A
影响因子：
56.9
作者：
Gamble, TR;Yoo, SH;Hill, CP
通讯作者：
Hill, CP
HIV 核衣壳蛋白。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
You,JC;McHenry,CS
通讯作者：
McHenry,CS
逆转录病毒结构蛋白与单链核酸的相互作用。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Karpel,RL;Henderson,LE;Oroszlan,S
通讯作者：
Oroszlan,S