Novel Nuclease MbovP701 with a Yqaj Domain Is Interrelated with the Growth of <i>Mycoplasma bovis</i>

<i>Mycoplasma bovis</i> (<i>M. bovis</i>) is characterized by a reduced genomic size and limited synthetic capacity, including the inability to synthesize nucleotides de novo, relies on nucleases for nutrient acquisition and survival. A number of nucleases have been implicate...

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Main Authors: Zhiyu Hao, Doukun Lu, Xixi Li, Abdul Raheem, Gang Zhao, Ali Sobhy Dawood, Yingyu Chen, Xi Chen, Changmin Hu, Jianguo Chen, Lei Zhang, Xifang Zhu, Aizhen Guo
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/12/12/2509
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Summary:<i>Mycoplasma bovis</i> (<i>M. bovis</i>) is characterized by a reduced genomic size and limited synthetic capacity, including the inability to synthesize nucleotides de novo, relies on nucleases for nutrient acquisition and survival. A number of nucleases have been implicated in <i>M. bovis</i> pathogenicity, facilitating substrate degradation and contributing to DNA repair mechanisms that enhance bacterial persistence. The present study confirmed that the T5.808 mutant, in which a novel nuclease gene (Mbov_0701) was disrupted by the mini-Tn4001 transposon, exhibits a growth defect when co-cultured with EBL cells. However, the restoration of Mbov_0701 resulted in the resumption of growth in the mutant. The characterization of MbovP701 revealed that it had high activity in hydrolyzing dsDNA with 5′- to 3′- polarity. Furthermore, the substrates of MbovP701 were extended to include linear dsDNA, ssDNA, RNA, and plasmid DNA. The exonuclease activity is dependent on the presence of Mn<sup>2+</sup> and/or Mg<sup>2+</sup> ions, with an optimal pH and temperature of 8.3 and 43 °C, respectively. The truncation experiments of rMbovP701 revealed that YqaJ (41–185 aa) is the key functional domain of MbovP701 exonuclease. In conclusion, the present study identified a novel nuclease in <i>M. bovis</i> that plays an essential role in the proliferation of this minimal organism. This finding elucidates the survival strategy and pathogenesis of <i>M. bovis</i>, suggesting a potential therapeutic strategy for the treatment of <i>M. bovis</i> through targeting the inhibition of MbovP701. Moreover, it provides a foundation for future investigations into the interactions between MbovP701 and other nucleases involved in <i>M. bovis</i> biology.
ISSN:2076-2607