Random insertion transposon mutagenesis of Mycobacterium fortuitum identified mutant defective in biofilm formation

Poonam Katoch; Kinam Gupta; Ragothaman M Yennamalli; Jitendraa Vashistt; Gopal Singh Bisht; Rahul Shrivastava

doi:10.1016/j.bbrc.2019.11.021

Random insertion transposon mutagenesis of Mycobacterium fortuitum identified mutant defective in biofilm formation

Biochem Biophys Res Commun. 2020 Jan 22;521(4):991-996. doi: 10.1016/j.bbrc.2019.11.021. Epub 2019 Nov 12.

Authors

Poonam Katoch¹, Kinam Gupta², Ragothaman M Yennamalli³, Jitendraa Vashistt⁴, Gopal Singh Bisht⁵, Rahul Shrivastava⁶

Affiliations

¹ Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173234, H.P., India. Electronic address: [email protected].
² Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173234, H.P., India; German Cancer Research Centre, DKFZ and Faculty of Biosciences, Heidelberg University, Heidelberg, Germany. Electronic address: [email protected].
³ Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173234, H.P., India; Translational Genomics and Immunomics Unit, VivagenDx Labs, Chennai 600042, India. Electronic address: [email protected].
⁴ Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173234, H.P., India. Electronic address: [email protected].
⁵ Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173234, H.P., India. Electronic address: [email protected].
⁶ Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173234, H.P., India. Electronic address: [email protected].

PMID: 31727369
DOI: 10.1016/j.bbrc.2019.11.021

Abstract

Mycobacterium fortuitum has emerged as a nosocomial infectious agent and biofilm formation attributed for the presence of this bacterium in hospital environment. Transposon random mutagenesis was used to identify membrane-proteins for biofilm formation in M. fortuitum. Ten mutants were shortlisted from a library of 450 mutants for examine their biofilm forming ability. Comparative biofilm ability with respect to wild type M. fortuitum ATCC 6841 showed an altered and delayed biofilm formation in one mutant namely, MT721. Sequence analysis revealed mutation in anthranilate phosphoribosyl transferase (MftrpD), which is associated with tryptophan operon. Functional interaction study of TrpD protein through STRING showed its interaction with chorismate utilizing proteins, majorly involved in synthesis of aromatic amino acid and folic acid, suggesting that biofilm establishment and maintenance requires components of central metabolism. Our study indicates important role of MftrpD in establishment and maintenance of biofilm by M. fortuitum, which may further be explored for drug discovery studies against mycobacterial infections.

Keywords: Anthranilate phosphoribosyltransferase; Biofilm; Mycobacterium fortuitum; Transposon mutagenesis; trpD.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Anthranilate Phosphoribosyltransferase / chemistry
Anthranilate Phosphoribosyltransferase / metabolism
Bacterial Proteins / chemistry
Bacterial Proteins / metabolism
Biofilms / growth & development*
Chorismic Acid / metabolism
DNA Transposable Elements / genetics*
Mutagenesis, Insertional / genetics*
Mutation / genetics*
Mycobacterium fortuitum / genetics*
Mycobacterium fortuitum / physiology*
Protein Interaction Mapping
Protein Structure, Secondary

Substances

Bacterial Proteins
DNA Transposable Elements
Anthranilate Phosphoribosyltransferase
Chorismic Acid