Effect of Phosphoglucosamine Mutase on Biofilm Formation and Antimicrobial Susceptibilities in M. Smegmatis GlmM Gene Knockdown Strain

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2013 Apr 18

PMID 23593488

Citations 10

Authors

Jian Kang

Liming Xu

Shufeng Yang

Wendan Yu

Shuo Liu

Yi Xin

Yufang Ma

Affiliations

Soon will be listed here.

Abstract

UDP-N-acetylglucosamine (UDP-GlcNAc) is a direct glycosyl donor of linker unit (L-Rhamnose-D-GlcNAc) and an essential precursor of peptidoglycan in mycobacteria. Phosphoglucosamine mutase (GlmM) is involved in the formation of glucosamine-1-phosphate from glucosamine-6-phosphate, the second step in UDP-GlcNAc biosynthetic pathway. We have demonstrated that GlmM protein is essential for the growth of M. smegmatis. To facilitate the analysis of the GlmM protein function in mycobacteria, a tetracycline inducible M. smegmatis glmM gene knockdown strain was constructed by using an antisense RNA technology. After induction with 20 ng/ml tetracycline, the expression of GlmM protein in glmM gene knockdown strain was significantly decreased, resulting in a decline of cell growth. The morphological changes of glmM gene knockdown strain induced with 20 ng/ml tetracycline have been observed by scanning electron microscope and transmission electron microscope. Furthermore, insufficient GlmM protein reduced the biofilm formation and increased the sensitivity to isoniazid and ethambutol in M. smegmatis, indicating that GlmM protein had effect on the biofilm formation and the senstivity to some anti-tuberculosis drugs targeting the cell wall. These results provide a new insight on GlmM functions in mycobacteria, suggesting that GlmM could be a potential target for development of new anti-tuberculosis drug.

Citing Articles

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References

Gandhi N, Moll A, Sturm A, Pawinski R, Govender T, Lalloo U . Extensively drug-resistant tuberculosis as a cause of death in patients co-infected with tuberculosis and HIV in a rural area of South Africa. Lancet. 2006; 368(9547):1575-80. DOI: 10.1016/S0140-6736(06)69573-1. View

Sassetti C, Boyd D, Rubin E . Genes required for mycobacterial growth defined by high density mutagenesis. Mol Microbiol. 2003; 48(1):77-84. DOI: 10.1046/j.1365-2958.2003.03425.x. View

Palomino J, Martin A, Camacho M, Guerra H, Swings J, Portaels F . Resazurin microtiter assay plate: simple and inexpensive method for detection of drug resistance in Mycobacterium tuberculosis. Antimicrob Agents Chemother. 2002; 46(8):2720-2. PMC: 127336. DOI: 10.1128/AAC.46.8.2720-2722.2002. View

Recht J, Martinez A, Torello S, Kolter R . Genetic analysis of sliding motility in Mycobacterium smegmatis. J Bacteriol. 2000; 182(15):4348-51. PMC: 101957. DOI: 10.1128/JB.182.15.4348-4351.2000. View

Zhang W, Jones V, Scherman M, Mahapatra S, Crick D, Bhamidi S . Expression, essentiality, and a microtiter plate assay for mycobacterial GlmU, the bifunctional glucosamine-1-phosphate acetyltransferase and N-acetylglucosamine-1-phosphate uridyltransferase. Int J Biochem Cell Biol. 2008; 40(11):2560-71. PMC: 2602953. DOI: 10.1016/j.biocel.2008.05.003. View

van Heijenoort J . Identification of the glmU gene encoding N-acetylglucosamine-1-phosphate uridyltransferase in Escherichia coli. J Bacteriol. 1993; 175(19):6150-7. PMC: 206709. DOI: 10.1128/jb.175.19.6150-6157.1993. View

Klein D, Ferre-DAmare A . Structural basis of glmS ribozyme activation by glucosamine-6-phosphate. Science. 2006; 313(5794):1752-6. DOI: 10.1126/science.1129666. View

Shimazu K, Takahashi Y, Uchikawa Y, Shimazu Y, Yajima A, Takashima E . Identification of the Streptococcus gordonii glmM gene encoding phosphoglucosamine mutase and its role in bacterial cell morphology, biofilm formation, and sensitivity to antibiotics. FEMS Immunol Med Microbiol. 2008; 53(2):166-77. DOI: 10.1111/j.1574-695X.2008.00410.x. View

Raetz C . Molecular genetics of membrane phospholipid synthesis. Annu Rev Genet. 1986; 20:253-95. DOI: 10.1146/annurev.ge.20.120186.001345. View

10.

Thomason M, Storz G . Bacterial antisense RNAs: how many are there, and what are they doing?. Annu Rev Genet. 2010; 44:167-88. PMC: 3030471. DOI: 10.1146/annurev-genet-102209-163523. View

11.

Ojha A, Anand M, Bhatt A, Kremer L, Jacobs Jr W, Hatfull G . GroEL1: a dedicated chaperone involved in mycolic acid biosynthesis during biofilm formation in mycobacteria. Cell. 2005; 123(5):861-73. DOI: 10.1016/j.cell.2005.09.012. View

12.

Brennan P . Structure, function, and biogenesis of the cell wall of Mycobacterium tuberculosis. Tuberculosis (Edinb). 2003; 83(1-3):91-7. DOI: 10.1016/s1472-9792(02)00089-6. View

13.

Jolly L, Wu S, van Heijenoort J, de Lencastre H, Tomasz A . The femR315 gene from Staphylococcus aureus, the interruption of which results in reduced methicillin resistance, encodes a phosphoglucosamine mutase. J Bacteriol. 1997; 179(17):5321-5. PMC: 179399. DOI: 10.1128/jb.179.17.5321-5325.1997. View

14.

Blokpoel M, Murphy H, OToole R, Wiles S, Runn E, Stewart G . Tetracycline-inducible gene regulation in mycobacteria. Nucleic Acids Res. 2005; 33(2):e22. PMC: 548381. DOI: 10.1093/nar/gni023. View

15.

Chen W, Kuo T . A simple and rapid method for the preparation of gram-negative bacterial genomic DNA. Nucleic Acids Res. 1993; 21(9):2260. PMC: 309503. DOI: 10.1093/nar/21.9.2260. View

16.

Qu H, Xin Y, Dong X, Ma Y . An rmlA gene encoding d-glucose-1-phosphate thymidylyltransferase is essential for mycobacterial growth. FEMS Microbiol Lett. 2007; 275(2):237-43. DOI: 10.1111/j.1574-6968.2007.00890.x. View

17.

van Heijenoort J . Characterization of the essential gene glmM encoding phosphoglucosamine mutase in Escherichia coli. J Biol Chem. 1996; 271(1):32-9. DOI: 10.1074/jbc.271.1.32. View

18.

Zhang Y, Mitchison D . The curious characteristics of pyrazinamide: a review. Int J Tuberc Lung Dis. 2003; 7(1):6-21. View

19.

Li S, Kang J, Yu W, Zhou Y, Zhang W, Xin Y . Identification of M. tuberculosis Rv3441c and M. smegmatis MSMEG_1556 and essentiality of M. smegmatis MSMEG_1556. PLoS One. 2012; 7(8):e42769. PMC: 3414508. DOI: 10.1371/journal.pone.0042769. View

20.

Dye C, Espinal M, Watt C, Mbiaga C, Williams B . Worldwide incidence of multidrug-resistant tuberculosis. J Infect Dis. 2002; 185(8):1197-202. DOI: 10.1086/339818. View