Role of fruA and csgA genes in gene expression during development of Myxococcus xanthus. Analysis by two-dimensional gel electrophoresis.
Two genes, fruA and csgA, encoding a putative transcription factor and C-factor, respectively, are essential for fruiting body formation of Myxococcus xanthus. To investigate the role of fruA and csgA genes in developmental gene expression, developing cells as well as vegetative cells of M. xanthus wild-type, fruA::Tc, and csgA731 strains were pulse-labeled with [(35)S]methionine, and the whole cell proteins were analyzed using two-dimensional immobilized pH gradient/SDS-PAGE. Differences in protein synthesis patterns among more than 700 protein spots were detected during development of the three strains. Fourteen proteins showing distinctly different expression patterns in mutant cells were analyzed in more detail. Five of the 14 proteins were identified as elongation factor Tu (EF-Tu), Dru, DofA, FruA, and protein S by immunoblot analysis and mass spectroscopy. A gene encoding DofA was cloned and sequenced. Although both fruA and csgA genes regulate early development of M. xanthus, they were found to differently regulate expression of several developmental genes. The production of six proteins, including DofA and protein S, was dependent on fruA, whereas the production of two proteins was dependent on csgA, and one protein was dependent on both fruA and csgA. To explain the present findings, a new model was presented in which different levels of FruA phosphorylation may distinctively regulate the expression of two groups of developmental genes.
Horiuchi
Takayuki
T
Department of Biology, Tokyo Metropolitan University, Minamiohsawa, Hachioji, Tokyo 192-0397, Japan.
Taoka
Masato
M
Isobe
Toshiaki
T
Komano
Teruya
T
Inouye
Sumiko
S
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D013058:Mass Spectrometry; D008954:Models, Biological; D008969:Molecular Sequence Data; D016941:Myxococcus xanthus / Q000235:genetics* / Q000502:physiology; D010455:Peptides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014157:Transcription Factors / Q000502:physiology*; D014357:Trypsin / Q000494:pharmacology
D001426:Bacterial Proteins; C063576:CsgA protein, Myxococcus xanthus; C104848:FruA protein, Myxococcus xanthus; D010455:Peptides; D014157:Transcription Factors; D014357:Trypsin