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解脲脲原体 OMC-P162 株的 II 型限制修饰系统。

Type II restriction modification system in Ureaplasma parvum OMC-P162 strain.

机构信息

Department of Developmental Medicine, Research Institute, Osaka Women's and Children's Hospital, Izumi City, Osaka, Japan.

College of Medicine, University of the Philippines Manila, Manila, Philippines.

出版信息

PLoS One. 2018 Oct 16;13(10):e0205328. doi: 10.1371/journal.pone.0205328. eCollection 2018.

DOI:10.1371/journal.pone.0205328
PMID:30325937
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6191088/
Abstract

Ureaplasma parvum serovar 3 strain, OMC-P162, was isolated from the human placenta of a preterm delivery at 26 weeks' gestation. In this study, we sequenced the complete genome of OMC-P162 and compared it with other serovar 3 strains isolated from patients with different clinical conditions. Ten unique genes in OMC-P162, five of which encoded for hypothetical proteins, were identified. Of these, genes UPV_229 and UPV_230 formed an operon whose open reading frames were predicted to code for a DNA methyltransferase and a hypothetical protein, respectively. DNA modification analysis of the OMC-P162 genome identified N4-methylcytosine (m4C) and N6-methyladenine (m6A), but not 5-methylocytosine (m5C). UPV230 recombinant protein displayed endonuclease activity and recognized the CATG sequence, resulting in a blunt cut between A and T. This restriction enzyme activity was identical to that of the cultivated OMC-P162 strain, suggesting that this restriction enzyme was naturally expressed in OMC-P162. We designated this enzyme as UpaP162. Treatment of pT7Blue plasmid with recombinant protein UPV229 completely blocked UpaP162 restriction enzyme activity. These results suggest that the UPV_229 and UPV_230 genes act as a type II restriction-modification system in Ureaplasma OMC-P162.

摘要

微小脲原体 3 血清型菌株 OMC-P162 从 26 周早产的人类胎盘中分离得到。在这项研究中，我们对 OMC-P162 的全基因组进行了测序，并与其他来自不同临床条件患者的 3 血清型菌株进行了比较。在 OMC-P162 中发现了 10 个独特的基因，其中 5 个编码假设蛋白。其中，基因 UPV_229 和 UPV_230 形成一个操纵子，其开放阅读框分别预测编码 DNA 甲基转移酶和假设蛋白。对 OMC-P162 基因组的 DNA 修饰分析鉴定出 N4-甲基胞嘧啶（m4C）和 N6-甲基腺嘌呤（m6A），但没有 5-甲基胞嘧啶（m5C）。UPV230 重组蛋白显示内切酶活性，并识别 CATG 序列，导致 A 和 T 之间的钝切割。这种限制性内切酶活性与培养的 OMC-P162 菌株相同，表明该限制性内切酶在 OMC-P162 中自然表达。我们将这种酶命名为 UpaP162。用重组蛋白 UPV229 处理 pT7Blue 质粒完全阻断了 UpaP162 限制酶的活性。这些结果表明，UPV_229 和 UPV_230 基因在微小脲原体 OMC-P162 中充当 II 型限制修饰系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/46a7950caade/pone.0205328.g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/3a71131dafd6/pone.0205328.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/0f5bb66b7139/pone.0205328.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/e4786d31f42b/pone.0205328.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/16b29ccf2e5c/pone.0205328.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/117dd83c360d/pone.0205328.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/809ce75ab029/pone.0205328.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/46da6ad5b95c/pone.0205328.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/527f76de9918/pone.0205328.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/6c528fa155ec/pone.0205328.g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/46a7950caade/pone.0205328.g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/3a71131dafd6/pone.0205328.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/0f5bb66b7139/pone.0205328.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/e4786d31f42b/pone.0205328.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/16b29ccf2e5c/pone.0205328.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/117dd83c360d/pone.0205328.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/809ce75ab029/pone.0205328.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/46da6ad5b95c/pone.0205328.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/527f76de9918/pone.0205328.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/6c528fa155ec/pone.0205328.g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f90d/6191088/46a7950caade/pone.0205328.g010.jpg

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