title: Assembling of the Mycobacterium tuberculosis Cell Wall Core.
abstract: The unique cell wall of mycobacteria is essential to their viability and the target of many clinically used anti-tuberculosis drugs and inhibitors under development. Despite intensive efforts to identify the ligase(s) responsible for the covalent attachment of the two major heteropolysaccharides of the mycobacterial cell wall, arabinogalactan (AG) and peptidoglycan (PG), the enzyme or enzymes responsible have remained elusive. We here report on the identification of the two enzymes of Mycobacterium tuberculosis, CpsA1 (Rv3267) and CpsA2 (Rv3484), responsible for this function. CpsA1 and CpsA2 belong to the widespread LytR-Cps2A-Psr (LCP) family of enzymes that has been shown to catalyze a variety of glycopolymer transfer reactions in Gram-positive bacteria, including the attachment of wall teichoic acids to PG. Although individual cpsA1 and cpsA2 knock-outs of M. tuberculosis were readily obtained, the combined inactivation of both genes appears to be lethal. In the closely related microorganism Corynebacterium glutamicum, the ortholog of cpsA1 is the only gene involved in this function, and its conditional knockdown leads to dramatic changes in the cell wall composition and morphology of the bacteria due to extensive shedding of cell wall material in the culture medium as a result of defective attachment of AG to PG. This work marks an important step in our understanding of the biogenesis of the unique cell envelope of mycobacteria and opens new opportunities for drug development.
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682.
authors.forename: Anna E
authors.initials: AE
authors.lastname: Grzegorzewicz
authors.affiliation: the Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Energie Atomique (CEA), CNRS, Université Paris Sud, F-91198 Gif-sur-Yvette, France.
authors.forename: Célia
authors.initials: C
authors.lastname: de Sousa-d'Auria
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682.
authors.forename: Michael R
authors.initials: MR
authors.lastname: McNeil
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682.
authors.forename: Emilie
authors.initials: E
authors.lastname: Huc-Claustre
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682.
authors.forename: Victoria
authors.initials: V
authors.lastname: Jones
authors.affiliation: the Department of Chemistry and Biochemistry, University of Toledo, Toledo, Ohio 43606-3390.
authors.forename: Cécile
authors.initials: C
authors.lastname: Petit
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682.
authors.forename: Shiva Kumar
authors.initials: SK
authors.lastname: Angala
authors.affiliation: the Department of Biochemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Mlynská dolina CH-1, 84215 Bratislava, Slovakia.
authors.forename: Júlia
authors.initials: J
authors.lastname: Zemanová
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682, the Key Laboratory of Medical Molecular Virology of MOE & MOH, Institutes of Biomedical Sciences and Institute of Medical Microbiology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors.forename: Qinglan
authors.initials: Q
authors.lastname: Wang
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682.
authors.forename: Juan Manuel
authors.initials: JM
authors.lastname: Belardinelli
authors.affiliation: the Key Laboratory of Medical Molecular Virology of MOE & MOH, Institutes of Biomedical Sciences and Institute of Medical Microbiology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Gao
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682, the Institute of Microbial Chemistry (BIKAKEN), Kamiosaki, Shinagawa-ku, Tokyo 3-14-23, Japan, and.
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Ishizaki
authors.affiliation: the Department of Biochemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Mlynská dolina CH-1, 84215 Bratislava, Slovakia.
authors.forename: Katarína
authors.initials: K
authors.lastname: Mikušová
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Brennan
authors.affiliation: the Department of Chemistry and Biochemistry, University of Toledo, Toledo, Ohio 43606-3390.
authors.forename: Donald R
authors.initials: DR
authors.lastname: Ronning
authors.affiliation: the C-CINA Center for Imaging and NanoAnalytics, Biozentrum, University of Basel, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Chami
authors.affiliation: the Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Energie Atomique (CEA), CNRS, Université Paris Sud, F-91198 Gif-sur-Yvette, France, christine.houssin@u-psud.fr.
authors.forename: Christine
authors.initials: C
authors.lastname: Houssin
authors.affiliation: From the Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682, Mary.Jackson@colostate.edu.
authors.forename: Mary
authors.initials: M
authors.lastname: Jackson
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism*; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism; D005685:Galactans / Q000235:genetics / Q000378:metabolism*; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism*; D010457:Peptidoglycan / Q000235:genetics / Q000378:metabolism*; D013682:Teichoic Acids / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D005685:Galactans; D010457:Peptidoglycan; D013682:Teichoic Acids; C005653:arabinogalactan
keywords: Mycobacterium tuberculosis; arabinogalactan; cell wall; cryo-electron microscopy; ligase; outer membrane; peptidoglycan; polysaccharide