Katsy E I
Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov, 410015 Russia.
Genetika. 2002 Jan;38(1):22-32.
The sequencing data were analyzed for two regions of the 120-MDa plasmid (p120) of Azospirillum brasilense Sp245. The 2420-bp region I, which flanks the omegon insertion in the SK048 mutant defective in production of the polar flagellum (Fla-) and swarming (Swa-), was shown to contain a cluster of two open reading frames (orf) that possess properties of coding sequences (CDSs). The NtrA (sigma 54) boxes were found in their upstream regions. The products of orf1 and orf2 are 16.5 and 15.5 kDa in molecular weight and consist of 151 and 152 amino-acid residues, respectively. The PRF1 polypeptide was found to contain a region homologous to the cysteine- and glycine-rich zinc-binding domain of the DnaJ heat-shock protein. ORF2 showed a homology to Haemophilus ducreyi pilin, fragments of Streptomyces and Mycobacterium integral membrane proteins, and eukaryotic transcriptional regulators. The omegon proved to be inserted into orfX1/X2 which possibly has a deletion and shows a GC content untypical for A. brasilense genes. The deduced ORFX2 polypeptide is homologous to fragments of arsenite-translocating ATPase and signal-transducing histidine kinase of archaebacteria. Possible causes of the Fla-Swa- phenotype of the A. brasilense SK048 mutant are considered. One coding orf was identified in the 1194-bp region II located approximately 4 kb away from the omegon insertion. The N-terminal region of the deduced product of this partly sequenced orf was shown to contain a signal sequence typical for secreted proteins.
对巴西固氮螺菌Sp245的120-MDa质粒(p120)的两个区域进行了测序数据分析。2420 bp的区域I位于SK048突变体中ω片段插入位点的侧翼,该突变体缺乏极鞭毛(Fla-)和群体游动(Swa-)能力,结果表明该区域包含两个开放阅读框(orf)组成的簇,这些开放阅读框具有编码序列(CDS)的特性。在它们的上游区域发现了NtrA(σ54)框。orf1和orf2的产物分子量分别为16.5 kDa和15.5 kDa,分别由151和152个氨基酸残基组成。发现PRF1多肽含有与DnaJ热休克蛋白富含半胱氨酸和甘氨酸的锌结合结构域同源的区域。ORF2与杜氏嗜血杆菌菌毛蛋白、链霉菌和分枝杆菌整合膜蛋白片段以及真核转录调节因子具有同源性。结果证明ω片段插入到orfX1/X2中,该orf可能存在缺失,并且其GC含量对于巴西固氮螺菌基因来说不典型。推导的ORFX2多肽与古细菌的亚砷酸盐转运ATP酶和信号转导组氨酸激酶片段同源。考虑了巴西固氮螺菌SK048突变体Fla-Swa-表型的可能原因。在距离ω片段插入位点约4 kb的1194 bp区域II中鉴定出一个编码orf。该部分测序的orf推导产物的N端区域显示含有分泌蛋白典型的信号序列。