1887

Abstract

The gene of , which encodes lipoamide dehydrogenase (EC 1.8.1.4), has been cloned and characterized. The gene is present as a single copy in the yeast genome and is transcribed to give a polyadenylated mRNA species of approximately 2·0 kb. The synthesis of lipoamide dehydrogenase in yeast is subject to carbon catabolite repression since both the level of the transcript and the accumulation of the lipoamide dehydrogenase subunit polypeptide were greatly reduced in wild-type cells grown on glucose compared to those grown on a variety of non-fermentable carbon sources. Strains defective in but transformed with the gene on a high copy number vector exhibited elevated levels of the transcript as well as increased lipoamide dehydrogenase activity when grown on glycerol. Immunoblotting experiments confirmed that such transformants over-expressed lipoamide dehydrogenase protein. Transcription from the sequence on plasmid pGP1 still appeared to be subject to some catabolite repression despite the presence of multiple copies of the plasmid in the cell.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-133-4-925
1987-04-01
2022-07-03
Loading full text...

Full text loading...

/deliver/fulltext/micro/133/4/mic-133-4-925.html?itemId=/content/journal/micro/10.1099/00221287-133-4-925&mimeType=html&fmt=ahah

References

  1. Beach D., Durkacz B., Nurse P. 1982; Functionally homologous cell cycle control genes in budding and fission yeast. Nature, London 300:706–709
    [Google Scholar]
  2. Beggs J. D. 1978; Transformation of yeast by a replicating hybrid plasmid. Nature, London 275:104–108
    [Google Scholar]
  3. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  4. Broach J. R., Strathern J. N., Hicks J. B. 1979; Transformation in yeast: develoμment of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–133
    [Google Scholar]
  5. Dagert M., Ehrlich S. D. 1979; Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 6:23–28
    [Google Scholar]
  6. Dickinson J. R., Roy D. J., Dawes I. W. 1986; A mutation affecting lipoamide dehydrogenase, pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase activities in Saccharomyces cerevisiae. . Molecular and General Genetics 204:103–107
    [Google Scholar]
  7. Guest J. R., Rice D. W. 1984; Molecular genetic approaches to the study of E. coli flavoproteins. In Flavins and Flavoproteins pp 111–123 Bray R. C. Edited by Berlin: Walter de Gruyter;
    [Google Scholar]
  8. Holmes D. S., Quigley M. 1981; A rapid boiling method for the preparation of bacterial plasmids. Analytical Biochemistry 114:193–197
    [Google Scholar]
  9. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual Cold Spring Harbor, New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  10. Mortimer R. K., Hawthorne D. C. 1975; Genetic mapping in yeast. Methods in Cell Biology 11:221–233
    [Google Scholar]
  11. Murherjee B. B., Matthews J., Horney O. L., Reed L. J. 1965; Resolution and reconstitution of Escherichia coli a-ketoglutarate dehydrogenase complex. Journal of Biological Chemistry 240:2268–2269
    [Google Scholar]
  12. Nasmyth K. A., Reed S. I. 1980; Isolation of genes by complementation in yeast: molecular cloning of a cell cycle gene. Proceedings of the National Academy of Sciences of the United States of America 77:2119–2123
    [Google Scholar]
  13. Nasmyth K. A., Tatchell K. 1980; The structure of transposable yeast mating type loci. Cell 19:753–764
    [Google Scholar]
  14. Polakis E. S., Bartley W. 1965; Changes in enzyme activities in S. cerevisiae during aerobic growth on different carbon sources. Biochemical Journal 97:284–297
    [Google Scholar]
  15. Polakis E. S., Bartley W., Meek G. A. 1965; Changes in the activities of respiratory enzymes during the aerobic growth of yeasts on different carbon sources. Biochemical Journal 97:298–302
    [Google Scholar]
  16. Prentri P., Karch F., Iida S., Meyer J. 1981; The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication. Gene 14:289–299
    [Google Scholar]
  17. Reed L. J. 1974; Multienzyme complexes. Accounts of Chemical Research 7:40–46
    [Google Scholar]
  18. Sherman F., Lawrence C. W. 1974; Saccharo- myces. In Handbook of Genetics 1 pp 359–393 King R. C. Edited by New York: Plenum Press;
    [Google Scholar]
  19. Smith H. O. 1980; Recovery of DNA from gels. Methods in Enzymology 65:371–380
    [Google Scholar]
  20. Spencer M. E., Guest J. R. 1985; Transcriptional analysis of the sucAB, aceEF and Ipd genes of Escherichia coli. . Molecular and General Genetics 200:145–154
    [Google Scholar]
  21. Szerely E., Montgomery D. L. 1984; Glucose represses transcription of Saccharomyces cerevisiae nuclear genes that encode mitochondrial components. Molecular and Cellular Biology 4:939–946
    [Google Scholar]
  22. Thomas P. S. 1980; Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proceedings of the National Academy of Sciences of the United States of America 11:5201–5205
    [Google Scholar]
  23. Weir-Thompson E. M., Dawes I. W. 1984; Develoμmental changes in translatable RNA species associated with meiosis and spore formation in Saccharomyces cerevisiae. . Molecular and Cellular Biology 4:695–702
    [Google Scholar]
  24. Winston F., Chumley F., Finr J. R. 1983; Eviction and transplacement of mutant genes in yeast. Methods in Enzymology 101:211–228
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-133-4-925
Loading
/content/journal/micro/10.1099/00221287-133-4-925
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error