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Mycoplasma agalactiae MAG_5040 is a Mg2+-dependent, sugar-nonspecific SNase recognised by the host humoral response during natural infection

Cacciotto, Carla and Addis, Maria Filippa and Coradduzza, Elisabetta and Carcangiu, Laura and Nuvoli, Anna Maria and Tore, Gessica and Dore, Gian Mario and Pagnozzi, Daniela and Uzzau, Sergio and Chessa, Bernardo and Pittau, Marco and Alberti, Alberto (2013) Mycoplasma agalactiaeMAG_5040 is a Mg2+-dependent, sugar-nonspecific SNase recognised by the host humoral response during natural infection. PLoS One, Vol. 8 (2), e57775. eISSN 1932-6203. Article.

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DOI: 10.1371/journal.pone.0057775

Abstract

In this study the enzymatic activity of Mycoplasma agalactiae MAG_5040, a magnesium-dependent nuclease homologue to the staphylococcal SNase was characterized and its antigenicity during natural infections was established. A UGA corrected version of MAG_5040, lacking the region encoding the signal peptide, was expressed in Escherichia coli as a GST fusion protein. Recombinant GST-MAG_5040 exhibits nuclease activity similar to typical sugar-nonspecific endo- and exonucleases, with DNA as the preferred substrate and optimal activity in the presence of 20 mM MgCl2 at temperatures ranging from 37 to 45uC. According to in silico analyses, the position of the gene encoding MAG_5040 is consistently located upstream an ABC transporter, in most sequenced mycoplasmas belonging to the Mycoplasma hominis group. In M. agalactiae, MAG_5040 is transcribed in a polycistronic RNA together with the ABC transporter components and with MAG_5030, which is predicted to be a sugar solute binding protein by 3D modeling and homology search. In a natural model of sheep and goats infection, anti-MAG_5040 antibodies were detected up to 9 months post infection. Taking into account its enzymatic activity, MAG_5040 could play a key role in Mycoplasma agalactiae survival into the host, contributing to host pathogenicity. The identification of MAG_5040 opens new perspectives for the development of suitable tools for the control of contagious agalactia in small ruminants.

Item Type:Article
ID Code:8781
Status:Published
Refereed:Yes
Uncontrolled Keywords:Mycoplasma agalactiae, MAG_5040, Escherichia coli
Subjects:Area 07 - Scienze agrarie e veterinarie > VET/05 Malattie infettive degli animali domestici
Area 06 - Scienze mediche > MED/07 Microbiologia e microbiologia clinica
Divisions:002 Altri enti e centri di ricerca del Nord Sardegna > Porto Conte Ricerche, Alghero
001 Università di Sassari > 01-a Nuovi Dipartimenti dal 2012 > Scienze Biomediche
001 Università di Sassari > 01-a Nuovi Dipartimenti dal 2012 > Medicina Veterinaria
Publisher:Public Library of Science
eISSN:1932-6203
Copyright Holders:© 2013 Cacciotto et al.
Deposited On:11 Mar 2013 17:18

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