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Int. J. Mol. Sci. 2017, 18(8), 1732; doi:10.3390/ijms18081732

TrmFO, a Fibronectin-Binding Adhesin of Mycoplasma bovis

1
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
2
The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
3
Hubei International Scientific and Technological Cooperation Base of Veterinary Epidemiology, Huazhong Agricultural University, Wuhan 430070, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 16 April 2017 / Revised: 24 July 2017 / Accepted: 2 August 2017 / Published: 9 August 2017
(This article belongs to the Collection Proteins and Protein-Ligand Interactions)
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Abstract

Mycoplasma bovis is an important pathogenic mycoplasma, causing the cattle industry serious economic losses. Adhesion is a crucial step in the mycoplasmas’ infection and colonization process; fibronectin (Fn), an extracellular matrix glycoprotein, is a molecular bridge between the bacterial adhesins and host cell receptors. The present study was designed to characterize the Fn-binding ability of methylenetetrahydrofolate-tRNA-(uracil-5-)-methyltransferase (TrmFO) and its role in M. bovis cytoadherence. The trmFO (MBOV_RS00785) gene was cloned and expressed in E. coli BL21, and polyclonal antibodies against the recombinant TrmFO (rTrmFO) were raised in rabbits. Immunoblotting demonstrated that TrmFO was an immunogenic component, and the TrmFO expression was conserved in different M. bovis isolates. The mycoplasmacidal assay further showed that in the presence of complement, rabbit anti-recombinant TrmFO serum exhibited remarkable mycoplasmacidal efficacy. TrmFO was detected in both the M. bovis membrane and cytoplasm. By ligand dot blot and enzyme-linked immunosorbent assay (ELISA) binding assay, we found that rTrmFO bound Fn in a dose-dependent manner. Immunostaining visualized by confocal laser scanning microscopy showed that rTrmFO had capacity to adhere to the embryonic bovine lung (EBL) cells. In addition, the adhesion of M. bovis and rTrmFO to EBL cells could be inhibited by anti-rTrmFO antibodies. To the best of our knowledge, this is the first report to characterize the Fn-binding ability of TrmFO and its role in the bacterial adhesion to host cells. View Full-Text
Keywords: Mycoplasma bovis; TrmFO; fibronectin; adhesion Mycoplasma bovis; TrmFO; fibronectin; adhesion
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Guo, Y.; Zhu, H.; Wang, J.; Huang, J.; Khan, F.A.; Zhang, J.; Guo, A.; Chen, X. TrmFO, a Fibronectin-Binding Adhesin of Mycoplasma bovis. Int. J. Mol. Sci. 2017, 18, 1732.

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