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Genome analysis of the staphylococcal temperate phage DW2 and functional studies on the endolysin and tail hydrolase

機(jī)譯：金黃色葡萄球菌溫帶噬菌體DW2的基因組分析及溶血素和尾巴水解酶的功能研究

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摘要

This study describes the genome of temperate Siphoviridae phage DW2, which is routinely propagated on Staphylococcus aureus DPC5246. The 41941 bp genome revealed an open reading frame (ORF1) which has a high level of homology with members of the resolvase subfamily of site-specific serine recombinase, involved in chromosomal integration and excision. In contrast, the majority of staphylococcal phages reported to date encode tyrosine recombinases. Two putative genes encoded by phage DW2 (ORF15 and ORF24) were highly homologous to the NWMN0273 and NWMN0280 genes encoding virulence factors carried on the genome of ϕNM4, a prophage in the genome of S. aureus Newman. Phage DW2 also encodes proteins highly homologous to two well-characterized Staphylococcus aureus pathogenicity island derepressors encoded by the staphylococcal helper phage 80α indicating that it may similarly act as a helper phage for mobility of pathogenicity islands in S. aureus. This study also focused on the enzybiotic potential of phage DW2. The structure of the putative endolysin and tail hydrolase were investigated and used as the basis for a cloning strategy to create recombinant peptidoglycan hydrolyzing proteins. After overexpression in E. coli, four of these proteins (LysDW2, THDW2, CHAPE1-153, and CHAPE1-163) were demonstrated to have hydrolytic activity against peptidoglycan of S. aureus and thus represent novel candidates for exploitation as enzybiotics.
機(jī)譯：這項(xiàng)研究描述了溫帶Siphoviridae噬菌體DW2的基因組，該基因組通常在金黃色葡萄球菌DPC5246上繁殖。 41941 bp基因組揭示了一個(gè)開放閱讀框（ORF1），它與位點(diǎn)特異性絲氨酸重組酶分辨酶亞家族的成員具有高度的同源性，參與染色體整合和切除。相反，迄今報(bào)道的大多數(shù)葡萄球菌噬菌體編碼酪氨酸重組酶。噬菌體DW2編碼的兩個(gè)推定基因（ORF15和ORF24）與encodingNM4基因組中編碼毒力因子的NWMN0273和NWMN0280基因高度同源，, NM4是金黃色葡萄球菌紐曼基因組中的噬菌體。噬菌體DW2還編碼與兩個(gè)由葡萄球菌輔助噬菌體80α編碼的特征明確的金黃色葡萄球菌致病島去阻遏物高度同源的蛋白質(zhì)，表明它可以類似地充當(dāng)金黃色葡萄球菌致病島的遷移的輔助噬菌體。該研究還集中于噬菌體DW2的酶促潛力。研究了推定的細(xì)胞溶素和尾巴水解酶的結(jié)構(gòu)，并將其用作克隆策略以創(chuàng)建重組肽聚糖水解蛋白的基礎(chǔ)。在大腸桿菌中過表達(dá)后，這些蛋白中的四種（LysDW2，THDW2，CHAPE1-153和CHAPE1-163）被證明具有抗金黃色葡萄球菌肽聚糖的水解活性，因此代表了新的候選酶類。

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