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. 1994 Sep 11;22(18):3773–3778. doi: 10.1093/nar/22.18.3773

The topoisomerase I gene from Ustilago maydis: sequence, disruption and mutant phenotype.

D Gerhold 1, M Thiyagarajan 1, E B Kmiec 1
PMCID: PMC308361  PMID: 7937091

Abstract

The Ustilago maydis genomic TOP1 gene encoding DNA topoisomerase I was cloned by amplifying a gene fragment using the polymerase chain reaction, and using this fragment to search a genomic DNA library by hybridization. The predicted peptide sequence exhibited 30-40% identity to other eukaryotic TOP1 genes, yet differed in several features. First, an unusually long acidic region was identified near the amino terminus (28/29 residues are acidic), which resembles other nucleolar peptide motifs. Second, an atypical carboxy-terminal 'tail', absent in other TOP1 genes, followed the active site tyrosine residue. A top1 gene disruption mutant was constructed by replacing the genomic TOP1 gene, with a top1::HygR null allele. This mutant lost the abundant topoisomerase I activity evident in wild-type U.maydis, and displayed a subtle coloration phenotype evident during cell senescence.

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Selected References

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