Robert F. Geever

DNA sequence, organization and regulation of the qa gene cluster of Neurospora crassa

In Neurospora, five structural and two regulatory genes mediate the initial events in quinate/shikimate metabolism as a carbon source. These genes are clustered in an 18 x 10(3) base-pair region as a contiguous array. The qa genes are induced by quinic acid and are coordinately controlled at the transcriptional level by the positive and negative regulators, qa-1F and qa-1S, respectively. The DNA sequence of the entire qa gene cluster has been determined and transcripts for each gene have been mapped. The qa genes are transcribed in divergent pairs and two types of transcripts are associated with each gene: basal level transcripts that initiate mainly from upstream regions and are independent of qa regulatory gene control, and inducible transcripts that initiate downstream from basal transcripts and are dependent on qa-1F binding to a 16 base-pair sequence. We discuss how both types of transcription relate to the organization of the qa genes as a cluster and how this may impose constraints on gene dispersal.

Comparative studies of the quinic acid (qa) cluster in several Neurospora species with special emphasis on the qa-x-qa-2 intergenic region

SummaryThe organization of the quinic acid (qa) genes in Neurospora crassa has been compared to that in several other Neurospora species. This gene cluster was found to be highly conserved in all species examined. However, there are numberous restriction fragment length polymorphisms that distinguish the heterothallic and homothallic species. Catabolic dehydroquinase assays indicated that qa-2 gene expression in the homothallic species is subject to induction by quinic acid, as is the case in N. crassa. The qa-x-qa-2 intergenic region of the homothallic species N. africana was cloned and sequenced. Conserved qa activator (qa-1F) binding sites have been identified in this region. When the qa-x-qa-2 intergenic region of N. crassa was replaced with its N. africana counterpart, qa-2 gene expression was reduced; however repression by glucose appeared normal. Furthermore, the N. africana start site for qa-2 transcription (which differs from the N. crassa start site) was utilized in the transformant. The overall evidence suggests that a weakening of the −120 activator binding site in the qa-x-qa-2 intergenic region may be responsible for these differences.

Sequence and characterization of the met-7 gene of Neurospora crassa ☆

The proteins encoded by the met-7+ and met-3+ genes of Neurospora crassa are required to form a functional cystathionine-gamma-synthase (CGS). The met-7+ gene has been cloned by complementation of a met-7 mutant. The nucleotide sequence of the complementing DNA reveals the presence of a 542-amino acid open reading frame (ORF). Disruption of this ORF abolishes complementation of the met-7 mutation.