Characterization of the spoT gene of Escherichia coli

E. Sarubbi, K. E. Rudd, H. Xiao, K. Ikehara, M. Kalman, M. Cashel

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

The Escherichia coli spoT gene encodes a guanosine-3',5'-bispyrophosphate (ppGpp) 3'-pyrophosphohydrolase known to be responsible for cellular (ppGpp) degradation. The DNA sequence of the spoT region is presented. The spoT gene is deduced to be 702 codons long, with a probable UUG initiation codon, and a deduced mass of 79,342 daltons. Two spoT mutations (spoT202 and spoT203) have been localized to an open reading frame by complementation of function as well as by genetic marker rescue. The ability to overexpress the spoT gene is limited, but enough ppGppase activity can be made to reverse ppGpp accumulation during the stringent response to amino acid starvation. The spoT gene is located within a larger spo operon and is flanked by two smaller genes. The first gene in the operon encodes omega, a protein that copurifies with RNA polymerase (Gentry, D.R., and Burgess, R.R. (1986) Gene (Amst.) 48, 33-40). The spoT gene is the second gene in the operon; it is followed by a third open reading frame deduced to encode a protein with a mass of 25,343 daltons. Insertion of a kanamycin resistance gene in the omega gene reduces spoT gene expression as judged by lowered ppGppase activity, relA-dependent reduction of growth rate, and abolition of spoT mutant complementation activity. These effects are reversed by expression of the spoT gene, but not the omega gene, in trans. Transcription of the spo operon occurs in a clockwise direction on the E. coli chromosome and is probably directed by at least two promoters.

Original languageEnglish
Pages (from-to)15074-15082
Number of pages9
JournalJournal of Biological Chemistry
Volume264
Issue number25
StatePublished - Jan 1 1989
Externally publishedYes

Fingerprint

Escherichia coli
Genes
Operon
Open Reading Frames
Kanamycin Resistance
Gene Expression
Initiator Codon
Guanosine
Kanamycin
DNA-Directed RNA Polymerases
DNA sequences
Starvation
Genetic Markers
Transcription
Codon
Chromosomes
Gene expression
Proteins
Amino Acids
Mutation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sarubbi, E., Rudd, K. E., Xiao, H., Ikehara, K., Kalman, M., & Cashel, M. (1989). Characterization of the spoT gene of Escherichia coli. Journal of Biological Chemistry, 264(25), 15074-15082.

Characterization of the spoT gene of Escherichia coli. / Sarubbi, E.; Rudd, K. E.; Xiao, H.; Ikehara, K.; Kalman, M.; Cashel, M.

In: Journal of Biological Chemistry, Vol. 264, No. 25, 01.01.1989, p. 15074-15082.

Research output: Contribution to journalArticle

Sarubbi, E, Rudd, KE, Xiao, H, Ikehara, K, Kalman, M & Cashel, M 1989, 'Characterization of the spoT gene of Escherichia coli', Journal of Biological Chemistry, vol. 264, no. 25, pp. 15074-15082.
Sarubbi E, Rudd KE, Xiao H, Ikehara K, Kalman M, Cashel M. Characterization of the spoT gene of Escherichia coli. Journal of Biological Chemistry. 1989 Jan 1;264(25):15074-15082.
Sarubbi, E. ; Rudd, K. E. ; Xiao, H. ; Ikehara, K. ; Kalman, M. ; Cashel, M. / Characterization of the spoT gene of Escherichia coli. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 25. pp. 15074-15082.
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N2 - The Escherichia coli spoT gene encodes a guanosine-3',5'-bispyrophosphate (ppGpp) 3'-pyrophosphohydrolase known to be responsible for cellular (ppGpp) degradation. The DNA sequence of the spoT region is presented. The spoT gene is deduced to be 702 codons long, with a probable UUG initiation codon, and a deduced mass of 79,342 daltons. Two spoT mutations (spoT202 and spoT203) have been localized to an open reading frame by complementation of function as well as by genetic marker rescue. The ability to overexpress the spoT gene is limited, but enough ppGppase activity can be made to reverse ppGpp accumulation during the stringent response to amino acid starvation. The spoT gene is located within a larger spo operon and is flanked by two smaller genes. The first gene in the operon encodes omega, a protein that copurifies with RNA polymerase (Gentry, D.R., and Burgess, R.R. (1986) Gene (Amst.) 48, 33-40). The spoT gene is the second gene in the operon; it is followed by a third open reading frame deduced to encode a protein with a mass of 25,343 daltons. Insertion of a kanamycin resistance gene in the omega gene reduces spoT gene expression as judged by lowered ppGppase activity, relA-dependent reduction of growth rate, and abolition of spoT mutant complementation activity. These effects are reversed by expression of the spoT gene, but not the omega gene, in trans. Transcription of the spo operon occurs in a clockwise direction on the E. coli chromosome and is probably directed by at least two promoters.

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