Department of Microbiology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
Proc Natl Acad Sci U S A. 2012 Mar 6;109(10):3962-6. doi: 10.1073/pnas.1119061109. Epub 2012 Feb 21.
Genetic variation is critical in microbial immune evasion and drug resistance, but variation has rarely been studied in complex heterogeneous communities such as the human microbiome. To begin to study natural variation, we analyzed DNA viruses present in the lower gastrointestinal tract of 12 human volunteers by determining 48 billion bases of viral DNA sequence. Viral genomes mostly showed low variation, but 51 loci of ∼100 bp showed extremely high variation, so that up to 96% of the viral genomes encoded unique amino acid sequences. Some hotspots of hypervariation were in genes homologous to the bacteriophage BPP-1 viral tail-fiber gene, which is known to be hypermutagenized by a unique reverse-transcriptase (RT)-based mechanism. Unexpectedly, other hypervariable loci in our data were in previously undescribed gene types, including genes encoding predicted Ig-superfamily proteins. Most of the hypervariable loci were linked to genes encoding RTs of a single clade, which we find is the most abundant clade among gut viruses but only a minor component of bacterial RT populations. Hypervariation was targeted to 5'-AAY-3' asparagine codons, which allows maximal chemical diversification of the encoded amino acids while avoiding formation of stop codons. These findings document widespread targeted hypervariation in the human gut virome, identify previously undescribed types of genes targeted for hypervariation, clarify association with RT gene clades, and motivate studies of hypervariation in the full human microbiome.
遗传变异在微生物的免疫逃避和耐药性方面至关重要,但在人类微生物组等复杂异质群落中,变异的研究很少。为了开始研究自然变异,我们通过测定 12 名人类志愿者下消化道中 480 亿个病毒 DNA 序列,分析了存在于其中的 DNA 病毒。病毒基因组大多显示出较低的变异,但有 51 个约 100 个碱基的位点显示出极高的变异,以至于多达 96%的病毒基因组编码了独特的氨基酸序列。一些超变异热点位于与噬菌体 BPP-1 病毒尾部纤维基因同源的基因中,已知该基因通过独特的逆转录酶(RT)机制发生超突变。出乎意料的是,我们数据中的其他超变位点位于以前未描述的基因类型中,包括编码预测的 Ig 超家族蛋白的基因。大多数高变位点与单一类群的 RT 基因相关,我们发现该类群是肠道病毒中最丰富的类群,但只是细菌 RT 群体的一小部分。超变针对 5'-AAY-3'天冬酰胺密码子,这允许编码的氨基酸最大程度的化学多样化,同时避免形成终止密码子。这些发现记录了人类肠道病毒组中广泛存在的靶向超变,确定了以前未描述的靶向超变的基因类型,阐明了与 RT 基因类群的关联,并激发了对完整人类微生物组中超变的研究。