The growth-promoting and stress response activities of the Bacillus subtilis GTP binding protein Obg are separable by mutation.
Bacillus subtilis Obg is a ribosome-associating GTP binding protein that is needed for growth, sporulation, and induction of the bacterium's general stress regulon (GSR). It is unclear whether the roles of Obg in sporulation and stress responsiveness are direct or a secondary effect of its growth-promoting functions. The present work addresses this question by an analysis of two obg alleles whose phenotypes argue for direct roles for Obg in each process. The first allele [obg(G92D)] encodes a missense change in the protein's highly conserved "obg fold" region. This mutation impairs cell growth and the ability of Obg to associate with ribosomes but fails to block sporulation or the induction of the GSR. The second obg mutation [obg(Delta22)] replaces the 22-amino-acid carboxy-terminal sequence of Obg with an alternative 26-amino-acid sequence. This Obg variant cofractionates with ribosomes and allows normal growth but blocks sporulation and impairs the induction of the GSR. Additional experiments revealed that the block on sporulation occurs early, preventing the activation of the essential sporulation transcription factor Spo0A, while inhibition of the GSR appears to involve a failure of the protein cascade that normally activates the GSR to effectively catalyze the reactions needed to activate the GSR transcription factor (sigma(B)).
Kuo
Shrin
S
Department of Microbiology and Immunology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA.
Demeler
Borries
B
Haldenwang
W G
WG
D000222:Adaptation, Physiological; D000595:Amino Acid Sequence; D019677:Artificial Gene Fusion; D001412:Bacillus subtilis / Q000235:genetics / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D008954:Models, Biological; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D011485:Protein Binding; D012270:Ribosomes / Q000378:metabolism; D017384:Sequence Deletion; D015398:Signal Transduction; D013171:Spores, Bacterial / Q000254:growth & development*; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D001426:Bacterial Proteins; C090542:Obg GTP-binding protein, Bacteria; C030069:spore-specific proteins, Bacillus; D001616:beta-Galactosidase; D019204:GTP-Binding Proteins