Molecular cloning and analysis of the regulation of cys-14/sup +/, a structural gene of the sulfur regulatory circuit of Neurospora crassa
The cys-14/sup +/ gene encodes sulfate permease II, which is primarily expressed in mycelia. cys-14/sup +/ is one of a set of sulfur-related structural genes under the control of cys-3/sup +/ and scon/sup +/, the regulatory genes of the sulfur control circuit. The authors cloned cys-14/sup +/ from a cosmid library of Neurospora crassa DNA. A restriction fragment length polymorphism analysis showed that this clone maps to the region of chromosome IV corresponding to the cys-14/sup +/ locus. Northern blot analyses were used to examine the regulated expression of the cys-14/sup +/ gene. In the wild type, a 3-kilobase cysc-14/sup +/ transcript was highly expressed under sulfur-derepressing conditions but completely absent during sulfur repression. A cys-3 mutant, which cannot synthesize any of the sulfur-controlled enzymes, including sulfate permease II, did not possess any cys-14/sup +/ transcript under wither condition. A cys-3 temperature-sensitive revertant completely lacked any cys-14/sup +/ mRNA at the conditional temperature but expressed the cys-14/sup +/ transcript upon derepression at the permissive temperature. Mutation of a second sulfur regulatory gene, scon/sup c/, causes the expression of sulfur-related enzymes in a constitutive fashion; the scon/sup c/ mutant showed a corresponding constitutive expression of cys-14/sup +/ mRNA, such that it was present even in cells subjected to sulfur repression conditions. These results show that the cys-14/sup +/ gene is regulated through the modulation of message content by the cys-3/sup +/ and scon/sup c/ control genes in response to the sulfur levels of the cells.
- Research Organization:
- The Ohio State Univ., Columbus, OH (US)
- OSTI ID:
- 6945045
- Journal Information:
- Mol. Cell. Biol.; (United States), Vol. 8:4
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ENZYMES
STRUCTURE-ACTIVITY RELATIONSHIPS
GENE RECOMBINATION PROTEINS
GENETIC MAPPING
NEUROSPORA
DNA-CLONING
GENE OPERONS
GENE REGULATION
MESSENGER-RNA
METABOLISM
MUTANTS
RFLPS
SULFUR
CLONING
ELEMENTS
FUNGI
MAPPING
NONMETALS
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
PLANTS
PROTEINS
RNA
550400* - Genetics
550200 - Biochemistry