MUTATIONAL ANALYSIS OF THE N-LINKED GLYCOSYLATION SITES OF THE SU ENVELOPE PROTEIN OF MOLONEY MURINE LEUKEMIA-VIRUS
MUTATIONAL ANALYSIS OF THE N-LINKED GLYCOSYLATION SITES OF THE SU ENVELOPE PROTEIN OF MOLONEY MURINE LEUKEMIA-VIRUS
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DOI：
10.1128/jvi.66.7.4258-4264.1992
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发表时间：
1992-07-01
影响因子：
5.4
通讯作者：
ROTH, MJ
中科院分区：
文献类型：
--
作者：
FELKNER, RH;ROTH, MJ
研究了Moloney小鼠白血病病毒主要包膜糖蛋白SU （gp70）中n -链糖基化位点的作用。通过使用位点特异性寡核苷酸定向诱变，七个聚糖添加位点中的每一个都被单独消除。突变导致单个糖基化位点的丢失，产生细胞内稳定的前体SU-TM蛋白，比野生型病毒SU-TM蛋白小4至5 kDa。突变体delta -1,4,7是一种缺乏三个n -链聚糖添加位点的营养物质，它产生了一个有活力的感染性病毒，其稳定的SU-TM蛋白比野生型SU-TM蛋白小约12至15 kDa。通过瞬时表达试验和XC合胞体试验的逆转录酶释放判断，7个单位点突变中有5个产生了活病毒。其中两个位点的突变导致了可检测的表型。2位突变的病毒在大鼠2细胞中对温度敏感；活病毒在32℃下产生，而在37℃下不产生。在3号位置突变的病毒没有传染性，产生的病毒粒子缺乏可检测到的成熟SU蛋白。突变导致蛋白质无法运输到细胞表面并组装成病毒粒子。
The role of the N-linked glycosylation sites in the major envelope glycoprotein, SU (gp70), of Moloney murine leukemia virus has been examined. By using site-specific oligonucleotide-directed mutagenesis, each of the seven glycan addition sites has been individually eliminated. Mutations resulting in the loss of a single glycosylation site produced, intracellularly, stable precursor SU-TM proteins which were 4 to 5 kDa smaller than the wild-type virus SU-TM protein. Mutant DELTA-1,4,7, a trimutant lacking three N-linked glycan addition sites, resulted in a viable, infectious virus with a stable SU-TM protein approximately 12 to 15 kDa smaller than the wild-type SU-TM protein. Five of the seven single-site mutations resulted in viable virus as judged by the release of reverse transcriptase in transient-expression assays and XC syncytium assays. Mutations at two of the sites resulted in a detectable phenotype. Virus mutated at position 2 was temperature sensitive in Rat2 cells; viable virus was produced at 32-degrees-C but not at 37-degrees-C. Virus mutated at position 3 was noninfectious and yielded virions lacking detectable mature SU protein. The mutation results in the block of transport of the protein to the cell surface and assembly into virion particles.