Characterization of the relA1 mutation and a comparison of relA1 with new relA null alleles in Escherichia coli.
The most widely studied "relaxed" mutant of the relA locus, the relA1 allele, is shown here to consist of an IS2 insertion between the 85th and 86th codons of the otherwise wild-type relA structural gene, which normally encodes a 743-amino acid (84 kDa) protein. The RelA protein is a ribosome-dependent ATP:GTP (GDP) pyrophosphoryltransferase that is activated during the stringent response to amino acid starvation and thereby occasions the accumulation of guanosine 3',5'-bispyrophosphate (ppGpp). We propose that the IS2 insertion functionally splits the RelA protein into two (alpha and beta) peptide fragments which can complement each other in trans to yield residual ppGpp synthetic activity; neither fragment shows this activity when expressed alone. Cell strains with a single copy relA null allele show physiological behavior that is much the same as relA1 mutant strains. Both relA1 and relA null strains accumulate ppGpp during glucose starvation and do not accumulate ppGpp during the stringent response. The presence of ppGpp in verifiable relA null strains is interpreted as unequivocal evidence for an alternate route of ppGpp synthesis that exists in addition to the relA-dependent reaction.
Metzger
S
S
Department of Cellular Biochemistry, Hebrew University Hadassah Medical School, Jerusalem, Israel.
Schreiber
G
G
Aizenman
E
E
Cashel
M
M
Glaser
G
G
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D001277:GTP Pyrophosphokinase / Q000235:genetics*; D005798:Genes, Bacterial; D005838:Genotype; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010770:Phosphotransferases / Q000235:genetics*; D010957:Plasmids; D015183:Restriction Mapping; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D010770:Phosphotransferases; D001277:GTP Pyrophosphokinase