Molecular cloning, expression, and characterization of a Ca2+-dependent, membrane-associated nuclease of Mycoplasma genitalium.

Journal of Bacteriology
Linbo LiThirumalai R Kannan

Abstract

In this study, we identified and characterized the enzymatic properties of MG_186, a calcium-dependent Mycoplasma genitalium nuclease. MG_186 displays the hallmarks of nucleases, as indicated by its amino acid sequence similarity to other nucleases. We cloned, UGA corrected, expressed, purified, and demonstrated that recombinant MG_186 (rMG_186) exhibits nuclease activity similar to that of typical sugar-nonspecific endonucleases and exonucleases. Biochemical characterization indicated that Ca2+ alone enhances its activity, which was inhibited by divalent cations, such as Zn2+ and Mn2+. Chelating agents EGTA and EDTA also inhibited nuclease activity. Mycoplasma membrane fractionation and Triton X-114 phase separation showed that MG_186 was a membrane-associated lipoprotein, and electron microscopy revealed its surface membrane location. Incubation of purified human endometrial cell nuclei with rMG_186 resulted in DNA degradation and morphological changes typical of apoptosis. Further, immunofluorescence analysis of rMG_186-treated nuclei indicated that morphological changes were linked to the disintegration of lamin and the internalization of rMG_186. Since M. genitalium has the capacity to invade eukaryotic cells and localize ...Continue Reading

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Citations

Jul 8, 2011·Clinical Microbiology Reviews·David Taylor-Robinson, Jørgen Skov Jensen
Jan 9, 2013·Infection and Immunity·Stephanie GroßhennigJörg Stülke
Oct 30, 2012·BMC Plant Biology·Krzysztof LeśniewiczPrzemysław Wojtaszek
Mar 5, 2016·Frontiers in Microbiology·Mitchell F Balish, Steven L Distelhorst
Mar 28, 2012·Protein Expression and Purification·Kunyuan GuoXinxin Zhang
Jan 24, 2015·Journal of Pathogens·Sunčanica Ljubin-Sternak, Tomislav Meštrović
May 3, 2016·International Journal of Molecular Sciences·Hui ZhangAizhen Guo
Feb 23, 2020·Applied Microbiology and Biotechnology·Lynn Sophie SchwardmannSkander Elleuche
Sep 10, 2019·Frontiers in Microbiology·Lianmei QinXiaoxing You
Dec 19, 2019·Frontiers in Microbiology·Xifang ZhuEric Baranowski
Apr 20, 2018·Frontiers in Cellular and Infection Microbiology·Peng LiWen-Xue Wu
Aug 11, 2021·Journal of Oncology·Elhem YacoubBoutheina Ben Abdelmoumen Mardassi

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