The sulfate activation locus of Escherichia coli K12

Cloning, genetic, and enzymatic characterization

Thomas S. Leyh, J. C. Taylor, G. D. Markham

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

The sulfate activation locus of Escherichia coli K12 has been cloned by complementation. The genes and gene products of this locus have been characterized by correlating the enzyme activity, complementation patterns, and polypeptides associated with subclones of the cloned DNA. The enzymes of the sulfate activation pathway, ATP sulfurylase (ATP:sulfate adenylyltransferase, EC 2.7.7.4) and APS kinase (APT:adenosine-5'-phosphosulfate 3'-phosphotransferase, EC 2.7.1.25) have been overproduced ~100-fold. Overproduction of ATP sulfurylase requires the expression of both the cysD gene, encoding a 27-kDa polypeptide, and a previously unidentified gene, denoted cysN, which encodes a 62-kDa polypeptide. Purification of ATP sulfurylase to homogeneity reveals that the enzyme is composed of two types of subunits which are encoded by cysD and cysN. Insertion of a kanamycin resistance gene into plasmid or chromosomal cysN prevents sulfate activation and decreases expression of the downstream cysC gene. cysC appears to be the APS kinase structural gene and encodes a 21-kDa polypeptide. The genes are adjacent and are transcribed counterclockwise on the E. coli chromosome in the order cysDNC. cysN and cysC are within the same operon and cysDNC are not in an operon containing cysHIJ.

Original languageEnglish (US)
Pages (from-to)2409-2416
Number of pages8
JournalJournal of Biological Chemistry
Volume263
Issue number5
StatePublished - 1988
Externally publishedYes

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Escherichia coli K12
Cloning
Sulfate Adenylyltransferase
Escherichia coli
Sulfates
Organism Cloning
adenylylsulfate kinase
Genes
Chemical activation
Peptides
Operon
Adenosine Phosphosulfate
Kanamycin Resistance
Gene encoding
Kanamycin
Enzyme activity
Enzymes
Enzyme Activation
Chromosomes
Purification

ASJC Scopus subject areas

  • Biochemistry

Cite this

The sulfate activation locus of Escherichia coli K12 : Cloning, genetic, and enzymatic characterization. / Leyh, Thomas S.; Taylor, J. C.; Markham, G. D.

In: Journal of Biological Chemistry, Vol. 263, No. 5, 1988, p. 2409-2416.

Research output: Contribution to journalArticle

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