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Biochemical genetics of Neurospora nuclease I: Isolation and characterization of nuclease (nuc) mutants

Published online by Cambridge University Press:  14 April 2009

A. M. Forsthoefel  and
N. C. Mishra
A. M. Forsthoefel
Affiliation:
Department of Biology, University of South Carolina, Columbia, SC 29208, U.S.A.
N. C. Mishra*
Affiliation:
Department of Biology, University of South Carolina, Columbia, SC 29208, U.S.A.
*
* Dr N. C. Mishra, Department of Biology, University of South Carolina, Columbia, SC 29208, U.S.A.

Summary

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Isolation and characterization of five new nuclease (nuc) deficient mutants of Neurospora have been described. The new mutants are unable to utilize nucleic acids as the sole phosphorus source and possess growth characteristics similar to those nuc (nuc-1 and nuc-2) mutants described previously. Two new mutants (nuc-4 and nuc-5) were able to use RNA or predigested DNA (but not intact DNA) as phosphorus source and showed temperature sensitive growth at 37 °C. Based on the data from complementation and genetic analyses the five new nuc mutants (nuc-3, nuc-4, nuc-5, nuc-6 and nuc-7) were found nonallelic to each other and to previously described nuc (nuc-1 and nuc-2) mutants; the new nuc mutants mapped to the right of arg-12 on linkage group II. On biochemical analyses, these nuc mutants were found to possess a lower level of extracellular nucleases and alkaline phosphatase as compared to the wild type strain. The ds DNase activity of the new mutants was only about 2–12% of that of the wild type strain; thus, the low level of these extracellular enzymes in the nuc mutants causes their inability to utilize nucleic acids as the sole phosphorus source. Wild type levels of these enzymes were restored in the complementing heterokaryons capable of full growth on the DNA medium. Data from intercrosses, mutagen sensitivity and spontaneous mutation-frequency studies (as discussed in a subsequent paper) indicated the involvement of the nuc genes in DNA repair and recombination.

Type
Research Article
Information
Genetics Research , Volume 41 , Issue 3 , June 1983 , pp. 271 - 286
Copyright
Copyright © Cambridge University Press 1983

References

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