Molecular and General Genetics MGG

, Volume 185, Issue 1, pp 142–147 | Cite as

Genetic mapping of a mutation in Escherichia coli leading to a temperature-sensitive RNase D

  • Richard Zaniewski
  • Murray R. Deutscher


In order to determine the metabolic role of RNase D in Escherichia coli, we have attempted to isolate strains deficient in this enzyme. One strain containing a temperature-sensitive RNase D was found among a heavily mutagenized stock of strains temperature-sensitive for growth. Genetic mapping of the mutation responsible for the altered RNase D enabled us to define the rnd locus, at 39.5–40.0 min on the E. coli map, which apparently specifies the RNase D structural gene. Using a Tn10 insertion near the rnd locus, we constructed isogenic strains containing RNase D and RNase II mutations, alone or in combination. Although the original mutant isolate displayed temperature-sensitive growth, no growth phenotype was associated with the rnd mutation in wild type background, possibly because a substantial amount of RNase D remained in cells grown at 45° C. However, elucidation of the map position of the rnd locus should prove useful for the isolation of other mutant strains with lower levels of RNase D.


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  1. Adelberg EA, Mandel M, Chen GCC (1965) Optimal conditions for mutagenesis by N-methyl-N′-nitro-N-nitrosoguanidine in Escherichia coli K12. Biochem Biophys Res Commun 18:788–795Google Scholar
  2. Bachman BJ, Low KB (1980) Linkage Map of Escherichia coli K12, Edition 6. Microbiol Rev 44:1–56Google Scholar
  3. Cudny H, Deutscher MP (1980) Apparent involvement of ribonuclease D in the 3′ processing of tRNA precursors. Proc Natl Acad Sci USA 77:837–841Google Scholar
  4. Cudny H, Roy P, Deutscher MP (1981a) Alteration of Escherichia coli RNase D by infection with bacteriophage T4. Biochem Biophys Res Commun 98:337–345Google Scholar
  5. Cudny H, Zaniewski R, Deutscher MP (1981b) Escherichia coli RNase D, purification and structural characterization of a putative processing nuclease. J Biol Chem 256:5627–5632Google Scholar
  6. Cudny H, Zaniewski R, Deutscher MP (1981c) Escherichia coli RNase D, catalytic properties and substrate specificity. J Biol Chem 256:5633–5637Google Scholar
  7. Dickinson ES, Sunderam TK (1970) Chromosomal location of a gene defining nicotinamide deamidase in E. coli. J Bacteriol 101:1090–1091Google Scholar
  8. Foulds J, Hilderman RH, Deutscher MP (1974) Mapping of the locus of Escherichia coli tRNA nucleotidyltransferase. J Bacteriol 118:628–632Google Scholar
  9. Fradkin JE, Fraenkel DG (1971) 2-keto-3-deoxygluconate 6-phosphate aldolase mutants of Escherichia coli. J Bacteriol 108:1277–1283Google Scholar
  10. Ghosh RK, Deutscher MP (1978a) Purification of potential 3′ processing nucleases using synthetic tRNA precursors. Nucleic Acids Res 5:3831–3842Google Scholar
  11. Ghosh RK, Deutscher MP (1978b) Identification of an Escherichia coli nuclease acting on structurally altered transfer RNA molecules. J Biol Chem 253:997–1000Google Scholar
  12. Hillman JD, Fraenkel DG (1975) Glyceraldehyde 3-phosphate dehydrogenase mutants of Escherichia coli. J Bacteriol 122:1175–1179Google Scholar
  13. Huang M, Pittard J (1967) Genetic analysis of mutant strains of Escherichia coli requiring p-aminobenzoic acid for growth. J Bacteriol 93:1938–1942Google Scholar
  14. Low B (1973) Rapid mapping of conditional and auxotrophic mutations in E. coli K12. J Bacteriol 113:798–812Google Scholar
  15. Miller JH (1972) Experiments in molecular gentics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  16. Nikolaev N, Folsom V, Schlessinger D (1976) Escherichia coli mutants deficient in exoribonucleases. Biochem Biophys Res Commun 70:920–924Google Scholar
  17. Ono M, Kuwano M (1977) Genetic analysis of mutations affecting ribonuclease II in Escherichia coli. Mol Gen Genet 153:1–4Google Scholar
  18. White BJ, Hochhauser SJ, Cintron NM, Weiss M (1976) Genetic mapping of xthA, the structural gene for exonuclease III in Escherichia coli K12. Bacteriol 126:1082–1088Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • Richard Zaniewski
    • 1
  • Murray R. Deutscher
    • 1
  1. 1.Department of BiochemistryUniversity of Connecticut, Health CenterFarmingtonUSA

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