The Accessory SecA2 System of Mycobacteria Requires ATP Binding and the Canonical SecA1.
In bacteria, the majority of exported proteins are transported by the general Sec pathway from their site of synthesis in the cytoplasm across the cytoplasmic membrane. The essential SecA ATPase powers this Sec-mediated export. Mycobacteria possess two nonredundant SecA homologs: SecA1 and SecA2. In pathogenic Mycobacterium tuberculosis and the nonpathogenic model mycobacterium Mycobacterium smegmatis, SecA1 is essential for protein export and is the "housekeeping" SecA, whereas SecA2 is an accessory SecA that exports a specific subset of proteins. In M. tuberculosis the accessory SecA2 pathway plays a role in virulence. In this study, we uncovered basic properties of the mycobacterial SecA2 protein and its pathway for exporting select proteins. By constructing secA2 mutant alleles that encode proteins defective in ATP binding, we showed that ATP binding is required for SecA2 function. SecA2 mutant proteins unable to bind ATP were nonfunctional and dominant negative. By evaluating the subcellular distribution of each SecA, SecA1 was shown to be equally divided between cytosolic and cell envelope fractions, whereas SecA2 was predominantly localized to the cytosol. Finally, we showed that the canonical SecA1 has a role in the process of SecA2-dependent export. The accessory SecA2 export system is important to the physiology and virulence of mycobacteria. These studies help establish the mechanism of this new type of specialized protein export pathway.
Rigel
Nathan W
NW
Department of Microbiology and Immunology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599-7290, USA.
Gibbons
Henry S
HS
McCann
Jessica R
JR
McDonough
Justin A
JA
Kurtz
Sherry
S
Braunstein
Miriam
M
D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000502:physiology*; D000255:Adenosine Triphosphate / Q000737:chemistry*; D000483:Alleles; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000502:physiology*; D003600:Cytosol / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D009154:Mutation; D009161:Mycobacterium / Q000378:metabolism*; D010641:Phenotype; D011485:Protein Binding; D013347:Subcellular Fractions
D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C442122:SecA1 protein, Mycobacterium tuberculosis; C442123:SecA2 protein, Mycobacterium