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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 238 (1993), S. 225-233 
    ISSN: 1617-4623
    Keywords: Neurospora crassa ; DNA repair ; RAD18 ; uvs-2 ; Zinc finger motif
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A clone containing the DNA repair gene uvs-2 of Neurospora crassa was identified from a Neurospora genomic DNA library using the sib-selection method. Transformants were screened for resistance to methyl methane sulfonate (MMS). A DNA fragment that complements the uvs-2 mutation was subcloned by monitoring its ability to transform the uvs-2 mutant to MMS resistance. Deletion analysis of the cloned DNA indicated that the size of the uvs-2 gene is approximately 1.6 kb. The identity of the uvs-2 gene was verified by restriction fragment length polymorphism (RFLP) mapping. The sensitivity of the transformant to three different mutagens was similar to that of the wild-type strain. Nucleotide sequences of genomic DNA and cDNA of the uvs-2 gene indicated that it has an open reading frame (ORF) of 1572 by with a 69 by intron in the middle of the sequence. Two transcription initiation sites located around 73 by and 290 by upstream of the translation initiation codon were identified by primer extension experiments. Northern analysis revealed that the nature transcript of the uvs-2 gene was about 1.8 kb long. The uvs-2 gene ORF is deduced to encode a polypeptide of 501 amino acids with a molecular mass of 54 kDa. The proposed polypeptide has 25% identity to the RAD18 polypeptide of Saccharomyces cerevisiae and contains two unique zinc finger motifs for nucleic acid binding. Similarities between the phenotypes of the rad18 and uvs-2 mutants suggest that the uvs-2 gene encodes a protein which is involved in postreplication repair, rather than excision repair.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Laccase ; Induction ; Cycloheximide ; Transcriptional regulation ; Neurospora crassa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A protein synthesis inhibitor, cycloheximide, induces excretion of laccase in Neurospora crassa. The lah-1 mutation results in excretion of a large amount of laccase even in the absence of cycloheximide. Ten mutations were induced that suppress derepressed excretion of laccase in the lah-1 mutant. Of these, seven second-site mutations were found to confer a laccase-noninducible phenotype, and were classified into two different complementation groups. Four mutations define a locus designated lni-1, found to be closely linked to ylo-1 on linkage group VI. The other three mutations were mapped to second locus, designated lni-2, that lies between nic-3 and thi-3on linkage group VII. The lni-2 locus was shown to encode laccase by RFLP mapping of the DNA fragment encoding laccase and by transformation of the lni-2 mutant with plasmid pBLI carrying the laccase gene (the locus encoding laccase is hereafter described as lacc). All lacc mutants examined (whether mutagen-induced or inactivated by repeat-induced point mutation) appeared to exhibit no phenotypic deficiency during both asexual and sexual cycles, suggesting that the laccase gene is dispensable in N. crassa. Northern analysis of total cellular RNA from the four lni-1 mutants demonstrated that the lni-1 mutations abolish increased transcription of the laccase gene under inducing conditions. Consequently, the lni-1 locus is inferred to encode a trans-acting positive regulator required for transcriptional activation of the laccase gene in response to cycloheximide. Possible functions of the lah-1 gene are also described.
    Type of Medium: Electronic Resource
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