PorA Represents the Major Cell Wall Channel of the Gram-positive Bacterium Corynebacterium Glutamicum

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 2003 Aug 5

PMID 12896997

Citations 12

Authors

Noelia Costa-Riu

Andreas Burkovski

Reinhard Kramer

Roland Benz

Affiliations

Soon will be listed here.

Abstract

The cell wall of the gram-positive bacterium Corynebacterium glutamicum contains a channel (porin) for the passage of hydrophilic solutes. The channel-forming polypeptide PorA is a 45-amino-acid acidic polypeptide with an excess of four negatively charged amino acids, which is encoded by the 138-bp gene porA. porA was deleted from the chromosome of C.glutamicum wild-type strain ATCC 13032 to obtain mutant ATCC 13032deltaporA. Southern blot analysis demonstrated that porA was deleted. Lipid bilayer experiments revealed that PorA was not present in the cell wall of the mutant strain. Searches within the known chromosome of C. glutamicum by using National Center for Biotechnology Information BLAST and reverse transcription-PCR showed that no other PorA-like protein is encoded on the chromosome or is expressed in the deletion strain. The porA deletion strain exhibited slower growth and longer growth times than the C. glutamicum wild-type strain. Experiments with different antibiotics revealed that the susceptibility of the mutant strain was much lower than that of the wild-type C. glutamicum strain. The results presented here suggest that PorA represents a major hydrophilic pathway through the cell wall and that C. glutamicum contains cell wall channels which are not related to PorA.

Citing Articles

Untargeted metabolomics coupled with genomics in the study of sucrose and xylose metabolism in .

Smoktunowicz M, Wawrzyniak R, Jonca J, Waleron M, Waleron K Front Microbiol. 2024; 15:1323765.

PMID: 38812674 PMC: 11133636. DOI: 10.3389/fmicb.2024.1323765.

Factors Determining the Susceptibility of Bacteria to Antibacterial Photodynamic Inactivation.

Rapacka-Zdonczyk A, Wozniak A, Michalska K, Pieranski M, Ogonowska P, Grinholc M Front Med (Lausanne). 2021; 8:642609.

PMID: 34055830 PMC: 8149737. DOI: 10.3389/fmed.2021.642609.

Identification and characterization of smallest pore-forming protein in the cell wall of pathogenic Corynebacterium urealyticum DSM 7109.

Abdali N, Younas F, Mafakheri S, Pothula K, Kleinekathofer U, Tauch A BMC Biochem. 2018; 19(1):3.

PMID: 29743008 PMC: 5944148. DOI: 10.1186/s12858-018-0093-9.

Click-chemistry approach to study mycoloylated proteins: Evidence for PorB and PorC porins mycoloylation in Corynebacterium glutamicum.

Issa H, Huc-Claustre E, Reddad T, Bonade Bottino N, Tropis M, Houssin C PLoS One. 2017; 12(2):e0171955.

PMID: 28199365 PMC: 5310785. DOI: 10.1371/journal.pone.0171955.

Corynebacterium jeikeium jk0268 constitutes for the 40 amino acid long PorACj, which forms a homooligomeric and anion-selective cell wall channel.

Abdali N, Barth E, Norouzy A, Schulz R, Nau W, Kleinekathofer U PLoS One. 2013; 8(10):e75651.

PMID: 24116064 PMC: 3792995. DOI: 10.1371/journal.pone.0075651.

References

Brennan P, Nikaido H . The envelope of mycobacteria. Annu Rev Biochem. 1995; 64:29-63. DOI: 10.1146/annurev.bi.64.070195.000333. View

Goodfellow M, Collins M, Minnikin D . Thin-layer chromatographic analysis of mycolic acid and other long-chain components in whole-organism methanolysates of coryneform and related taxa. J Gen Microbiol. 1976; 96(2):351-8. DOI: 10.1099/00221287-96-2-351. View

Lichtinger T, Burkovski A, Niederweis M, Kramer R, Benz R . Biochemical and biophysical characterization of the cell wall porin of Corynebacterium glutamicum: the channel is formed by a low molecular mass polypeptide. Biochemistry. 1998; 37(43):15024-32. DOI: 10.1021/bi980961e. View

Yano I, Saito K . Gas chromatographic and mass spectrometric analysis of molecular species of corynomycolic acids from Corynebacterium ulcerans. FEBS Lett. 1972; 23(3):352-356. DOI: 10.1016/0014-5793(72)80314-4. View

Steinmetz M, Le Coq D, Djemia H, Gay P . [Genetic analysis of sacB, the structural gene of a secreted enzyme, levansucrase of Bacillus subtilis Marburg]. Mol Gen Genet. 1983; 191(1):138-44. DOI: 10.1007/BF00330901. View

Benz R, Janko K, Boos W, Lauger P . Formation of large, ion-permeable membrane channels by the matrix protein (porin) of Escherichia coli. Biochim Biophys Acta. 1978; 511(3):305-19. DOI: 10.1016/0005-2736(78)90269-9. View

Lichtinger T, Riess F, Burkovski A, Engelbrecht F, Hesse D, Kratzin H . The low-molecular-mass subunit of the cell wall channel of the Gram-positive Corynebacterium glutamicum. Immunological localization, cloning and sequencing of its gene porA. Eur J Biochem. 2001; 268(2):462-9. DOI: 10.1046/j.1432-1033.2001.01913.x. View

van der Rest M, Lange C, Molenaar D . A heat shock following electroporation induces highly efficient transformation of Corynebacterium glutamicum with xenogeneic plasmid DNA. Appl Microbiol Biotechnol. 1999; 52(4):541-5. DOI: 10.1007/s002530051557. View

Beckers G, Nolden L, Burkovski A . Glutamate synthase of Corynebacterium glutamicum is not essential for glutamate synthesis and is regulated by the nitrogen status. Microbiology (Reading). 2001; 147(Pt 11):2961-70. DOI: 10.1099/00221287-147-11-2961. View

10.

West I, Page M . When is the outer membrane of Escherichia coli rate-limiting for uptake of galactosides?. J Theor Biol. 1984; 110(1):11-9. DOI: 10.1016/s0022-5193(84)80011-9. View

11.

Ochi K . Phylogenetic analysis of mycolic acid-containing wall-chemotype IV actinomycetes and allied taxa by partial sequencing of ribosomal protein AT-L30. Int J Syst Bacteriol. 1995; 45(4):653-60. DOI: 10.1099/00207713-45-4-653. View

12.

Trias J, Benz R . Permeability of the cell wall of Mycobacterium smegmatis. Mol Microbiol. 1994; 14(2):283-90. DOI: 10.1111/j.1365-2958.1994.tb01289.x. View

13.

Udaka S . Screening method for microorganisms accumulating metabolites and its use in the isolation of Micrococcus glutamicus. J Bacteriol. 1960; 79:754-5. PMC: 278770. DOI: 10.1128/jb.79.5.754-755.1960. View

14.

Gutmann M, Hoischen C, Kramer R . Carrier-mediated glutamate secretion by Corynebacterium glutamicum under biotin limitation. Biochim Biophys Acta. 1992; 1112(1):115-23. DOI: 10.1016/0005-2736(92)90261-j. View

15.

Puech V, Chami M, Lemassu A, Laneelle M, Schiffler B, Gounon P . Structure of the cell envelope of corynebacteria: importance of the non-covalently bound lipids in the formation of the cell wall permeability barrier and fracture plane. Microbiology (Reading). 2001; 147(Pt 5):1365-1382. DOI: 10.1099/00221287-147-5-1365. View

16.

Saiki R, Chang C, Levenson C, Warren T, Boehm C, Kazazian Jr H . Diagnosis of sickle cell anemia and beta-thalassemia with enzymatically amplified DNA and nonradioactive allele-specific oligonucleotide probes. N Engl J Med. 1988; 319(9):537-41. DOI: 10.1056/NEJM198809013190903. View

17.

Jager W, Schafer A, Puhler A, Labes G, Wohlleben W . Expression of the Bacillus subtilis sacB gene leads to sucrose sensitivity in the gram-positive bacterium Corynebacterium glutamicum but not in Streptomyces lividans. J Bacteriol. 1992; 174(16):5462-5. PMC: 206388. DOI: 10.1128/jb.174.16.5462-5465.1992. View

18.

Niederweis M, Ehrt S, Heinz C, Klocker U, Karosi S, Swiderek K . Cloning of the mspA gene encoding a porin from Mycobacterium smegmatis. Mol Microbiol. 1999; 33(5):933-45. DOI: 10.1046/j.1365-2958.1999.01472.x. View

19.

Neuhoff V, Arold N, Taube D, Ehrhardt W . Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250. Electrophoresis. 1988; 9(6):255-62. DOI: 10.1002/elps.1150090603. View

20.

Liu J, Barry 3rd C, Besra G, Nikaido H . Mycolic acid structure determines the fluidity of the mycobacterial cell wall. J Biol Chem. 1996; 271(47):29545-51. DOI: 10.1074/jbc.271.47.29545. View