Phosphatidylinositol is an essential phospholipid of mycobacteria.
Phosphatidylinositol (PI) and metabolically derived products such as the phosphatidylinositol mannosides and linear and mature branched lipomannan and lipoarabinomannan are prominent phospholipids/lipoglycans of Mycobacterium sp. believed to play important roles in the structure and physiology of the bacterium as well as during host infection. To determine if PI is an essential phospholipid of mycobacteria, we identified the pgsA gene of Mycobacterium tuberculosis encoding the phosphatidylinositol synthase enzyme and constructed a pgsA conditional mutant of Mycobacterium smegmatis. The ability of this mutant to synthesize phosphatidylinositol synthase and subsequently PI was dependent on the presence of a functional copy of the pgsA gene carried on a thermosensitive plasmid. The mutant grew like the control strain under permissive conditions (30 degrees C), but ceased growing when placed at 42 degrees C, a temperature at which the rescue plasmid is lost. Loss of cell viability at 42 degrees C was observed when PI and phosphatidylinositol dimannoside contents dropped to approximately 30 and 50% of the wild-type levels, respectively. This work provides the first evidence of the essentiality of PI to the survival of mycobacteria. PI synthase is thus an essential enzyme of Mycobacterium that shows promise as a drug target for anti-tuberculosis therapy.
Jackson
M
M
Department of Microbiology, Colorado State University, Fort Collins, Colorado 80523, USA.
Crick
D C
DC
Brennan
P J
PJ
D000595:Amino Acid Sequence; D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase; D003434:Crossing Over, Genetic; D005798:Genes, Bacterial; D005804:Genes, Lethal; D008969:Molecular Sequence Data; D009154:Mutation; D009161:Mycobacterium / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D020102:Mycobacterium smegmatis / Q000235:genetics / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism*; D010743:Phospholipids / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics* / Q000378:metabolism
D010716:Phosphatidylinositols; D010743:Phospholipids; C054977:phosphatidylinositol dimannoside; D017855:Transferases (Other Substituted Phosphate Groups); D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase; C020676:CDP-diacylglycerol-glycerol-3-phosphate 3-phosphatidyltransferase