A sulfur- and tyramine-regulated Klebsiella aerogenes operon containing the arylsulfatase (atsA) gene and the atsB gene

Abstract

The structural gene for arylsulfatase (atsA) of Klebsiella aerogenes was cloned into a pKI212 vector in Escherichia coli. Deletion analysis showed that the atsA gene with the promoter region was located within a 3.2-kilobase cloned segment. In E. colh cells which carried the plasmid, the synthesis of arylsulfatase was repressed by various sources of sulfur; the repression was relieved, in each case, by tyramine. Transfer of the plasmid into atsA or constitutive atsR mutant strains of K. aerogenes resulted in complementation of atsA but not of atsR. The nucleotide sequence of the 3.2-kilobase fragment was determined. Two open reading frames, the atsA gene and an unknown gene (atsB), were found. These are located between a potential promoter and a transcriptional terminator sequence. Deletion analysis suggests that atsB is a potential positive factor for the regulation of arylsulfatase. Analysis of the amino acid sequences of the first 13 amino acids from the N terminus of the purified secreted arylsulfatase agrees with that of the nucleotide sequence of atsA. The leader peptide extends over 20 amino acids and has the characteristics of a signal sequence. Primer extension mapping of transcripts generated in vivo suggests that the synthesis of mRNA starts at a site 31 or 32 bases upstream from the ATG initiation codon of the atsB gene. By Northern (RNA) blot analysis of the transcripts induced by tyramine, we found a 2.7-kilobase transcript which is identical in size to the total sequence of the atsB and atsA genes. Thus, the ats operon is composed of two cistrons and is regulated by sulfur and tyramine.

A monoamine-regulated Klebsiella aerogenes operon containing the monoamine oxidase structural gene (maoA) and the maoC gene

“…The bacterial strains used in this study are listed in Table 1. pKI212 (18), a plasmid conferring resistance to kanamycin (Kmr), and pTO58, a plasmid containing the maoA gene from K aerogenes W70, were constructed previously (28). pMC1403 (4), a plasmid containing the lacZ gene without the promoter region, was provided by A. Nakazawa (Yamaguchi University).…”

“…The nucleotide and amino acid se- (20 p,g) and primer extension with avian myeloblastosis virus reverse transcriptase were performed as described previously (18).…”

“…Coordinate expression of the maoA gene and the ats operon occurs (22). The ats operon is closely linked to maoA (15,17) and contains the genes for arylsulfatase (atsA) and the positive factor (atsB) for atsA synthesis (18). The F' episome from Escherichia coli and the transducing phage P1 were used to * Corresponding author.…”

Mutants of Chlamydomonas with Aberrant Responses to Sulfur Deprivation.

“…In K. aerogenes, transcription of ars is inhibited by sulfate or cysteine (Adachi et al, 1974;Murooka et al, 1990;Azakami et al, 1992). In N. crassa, the accumulation of Ars and the mycelial-specific permease is induced by growing mycelia in medium containing low levels of sulfur.…”

Sac3, an Snf1-like Serine/Threonine Kinase That Positively and Negatively Regulates the Responses of Chlamydomonas to Sulfur Limitation

“…However, the level of available inorganic sulfate in the soil may be low (David et al, 1982; Autry and Fitzgerald, 1990;Whalen and Warman, 1996). Many soil-dwelling microbes have developed mechanisms to scavenge sulfur from their environment by accessing organic sulfate esters, sulfamates, and sulfonates (Marzluf, 1970;Scott and Metzenberg, 1970; Apte et al, 1974;Lien and Schreiner, 1975; de Hostos et al, 1988;Murooka et al, 1990;Yildiz et al, 1994; Beil et al, 1996), which are abundant in the soil (Fitzgerald et al, 1988; Autry et al, 1990; Dhamala et al, 1990; Dhamala and Mitchell, 1995). In addition, both microbes and plants exhibit an increased efficiency for the import of sulfate when sulfate supplies are limiting (Breton and Surdin-Kerjan, 1977; Clarkson et al, 1983; Biedlingmaier and Schmidt, 1988; Green and Grossman, 1988;Hawkesford et al, 1993;Yildiz et al, 1994;Lappartient and Touraine, 1996).…”