The human cytomegalovirus UL133-138 gene locus attenuates the lytic viral cycle in fibroblasts.

The human cytomegalovirus UL133-138 gene locus attenuates the lytic viral cycle in fibroblasts.
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DOI:
10.1371/journal.pone.0120946
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发表时间:
2015
期刊:
影响因子:
3.7
通讯作者:
Stinski MF
Stinski MF
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Dutta N;Lashmit P;Yuan J;Meier J;Stinski MF

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HCMV临床株(如FIX、TR、PH等)的基因组包含一个15 kb的区域,编码20个推定的ORF。称为ULb '的区域在病毒在人包皮成纤维细胞(HFF)培养物中连续传代后丢失。与临床菌株相比,实验室菌株复制更快,感染性病毒滴度更高。我们使用FIX和TR作为模型临床菌株制备了具有从ULb'区域缺失的22、14或7个ORF的重组病毒。我们还在UL133和UL138之间的单个ORF中引入了终止密码子以阻止蛋白质表达。与野生型病毒相比,ULb'内的所有缺失和UL133至UL138区域内的所有终止密码子突变体、病毒主要立即早期RNA和蛋白质、DNA和无细胞感染性病毒不同程度地增加。野生型病毒蛋白质沿着无细胞感染性病毒从人成纤维细胞释放减慢病毒复制过程。
The genomes of HCMV clinical strains (e.g. FIX, TR, PH, etc) contain a 15 kb region that encodes 20 putative ORFs. The region, termed ULb’, is lost after serial passage of virus in human foreskin fibroblast (HFF) cell culture. Compared to clinical strains, laboratory strains replicate faster and to higher titers of infectious virus. We made recombinant viruses with 22, 14, or 7 ORFs deleted from the ULb’ region using FIX and TR as model clinical strains. We also introduced a stop codon into single ORFs between UL133 and UL138 to prevent protein expression. All deletions within ULb’ and all stop codon mutants within the UL133 to UL138 region increased to varying degrees, viral major immediate early RNA and protein, DNA, and cell-free infectious virus compared to the wild type viruses. The wild type viral proteins slowed down the viral replication process along with cell-free infectious virus release from human fibroblast cells.
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