Characterization of a Haloferax volcanii member of the enolase superfamily: deletion mutant construction, expression analysis, and transcriptome comparison.
The enolase superfamily (COG4948) contains proteins with very different biological functions including regulators like the Escherichia coli RspA and metabolic enzymes like enolase. To unravel the biological function of an archaeal family member, an in frame deletion mutant of a gene encoding a COG4948 protein of Haloferax volcanii was generated. The mutant had a lag phase of 3 days after transition from a richer to a poorer medium, in contrast to the wild-type, and the gene was therefore named "important for transition" (iftA). After inoculation of fresh casamino acids or complex medium with stationary phase wild-type cells, the transcript level of iftA was transiently induced at the onset of growth. In contrast, in minimal (or "poor") glucose medium, both transcript and protein were present throughout growth, even in late stationary phase. A comparison of the transcriptomes of deletion mutant and wild-type revealed that transcript levels of a very restricted set of genes were differentially regulated, including genes encoding proteins involved in phosphate metabolism, regulators and stress response proteins. Taken together, the results indicate that IftA might have a dual function, i.e., transiently after transition to fresh medium and permanently during growth in glucose medium.
Dambeck
Michael
M
Biocentre, Institute for Molecular Biosciences, Goethe-University, Max-von-Laue-Str. 9, 60438 Frankfurt, Germany. m.dambeck@bio.uni-frankfurt.de
Soppa
Jörg
J
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D019843:Archaeal Proteins / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D019848:Gene Expression Regulation, Archaeal; D019766:Genes, Archaeal; D019622:Haloferax volcanii / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000378:metabolism; D019642:RNA, Archaeal / Q000235:genetics; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D017384:Sequence Deletion
D000596:Amino Acids; D019843:Archaeal Proteins; D019642:RNA, Archaeal; C017721:casamino acids; D010751:Phosphopyruvate Hydratase