Previous examination revealed a correlation of phytopathogenic data of 'Candidatus Phytoplasma mali' strains and the DNA sequence variability of a type ATP00464 hflB gene fragment. To further investigate such a relationship, all distinct genes previously annotated as hflB in the genome of 'Ca. P. mali' strain AT were fully sequenced and analyzed from a number of representative mild, moderate, and severe strains. The re-annotation indicated that the sequences encode six AAA+ ATPases and six HflB proteases. Each of the nine distinct deduced AAA+ proteins that were examined formed a coherent phylogenetic cluster. However, within these groups, sequences of three ATPases and three proteases from mild and severe strains clustered distantly, according to their virulence. This grouping was supported by an association with virulence-related amino acid substitutions. Another finding was that full-length genes from ATPase AP11 could only be identified in mild and moderate strains. Prediction of the membrane topology indicated that the long ATPase- and protease-carrying C-terminal tails of approximately half of the AAA+ proteins are extracellular, putatively facing the environment of the sieve tubes. Thus, they may be involved in pathogen-host interactions and may compromise phloem function, a major effect of phytoplasma infection. All full-length genes examined appear transcriptionally active and all deduced peptides show the key positions indicative for protein function.

译文

先前的检查显示,“假丝酵母菌” 菌株的植物病原学数据与ATP00464型hflB基因片段的DNA序列变异性之间存在相关性。为了进一步研究这种关系,所有先前在 'Ca基因组中注释为hflB的不同基因。对P. mali' 菌株AT进行了完整的测序,并从许多代表性的轻度,中度和重度菌株中进行了分析。重新注释表明该序列编码六个AAA atpase和六个HflB蛋白酶。检查的九种不同的推导AAA蛋白中的每一种都形成了一个相干的系统发育簇。然而,在这些组中,根据其毒力,来自轻度和重度菌株的三种atpase和三种蛋白酶的序列远距聚集。该分组得到了与毒力相关氨基酸取代相关的关联的支持。另一个发现是,ATPase AP11的全长基因只能在轻度和中度菌株中鉴定。膜拓扑的预测表明,大约一半的AAA蛋白的长ATPase和蛋白酶携带的C末端尾巴是细胞外的,假定面对筛管的环境。因此,它们可能参与病原体与宿主的相互作用,并可能损害韧皮部功能,这是植原体感染的主要作用。所检查的所有全长基因均具有转录活性,并且所有推导的肽均显示出指示蛋白质功能的关键位置。

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