Abstract
The hexokinase A (HKA) and hexokinase B (HKB) genes of Saccharomyces cerevisiae have been cloned from a library of yeast genomic DNA. Using an in vitro glucose phosphorylation assay, the HKB gene was located on a plasmid carrying a 13.6 kb fragment of yeast DNA. After subcloning the relevant restriction fragments, the nucleotide sequence of the HKB gene was determined. Using this information, we were able to locate the HKA gene on a plasmid carrying this gene, which we then sequenced. Approximately 43% of the amino acid sequence of HKB was determined directly from 24 tryptic peptides. The results are in complete agreement with those derived from the DNA sequence and are consistent with the results of x-ray crystallography. Comparison of the amino acid sequences of HKA and HKB show that 378 out of 485 residues are identical. The 5' flanking region of the A gene contains nucleotide sequences expected for genes that are expressed at relatively high levels in yeast. The 24 base pair hyphenated palindrome at the 3' end of the HKB gene may be a site for termination of transcription of this gene.
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson C. M., McDonald R. C., Steitz T. A. Sequencing a protein by x-ray crystallography. I. Interpretation of yeast hexokinase B at 2.5 A resolution by model building. J Mol Biol. 1978 Jul 25;123(1):1–13. doi: 10.1016/0022-2836(78)90373-x. [DOI] [PubMed] [Google Scholar]
- Anderson C. M., Stenkamp R. E., Steitz T. A. Sequencing a protein by x-ray crystallography. II. Refinement of yeast hexokinase B co-ordinates and sequence at 2.1 A resolution. J Mol Biol. 1978 Jul 25;123(1):15–33. doi: 10.1016/0022-2836(78)90374-1. [DOI] [PubMed] [Google Scholar]
- Bennett W. S., Jr, Steitz T. A. Glucose-induced conformational change in yeast hexokinase. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4848–4852. doi: 10.1073/pnas.75.10.4848. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bennetzen J. L., Hall B. D. The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase. J Biol Chem. 1982 Mar 25;257(6):3018–3025. [PubMed] [Google Scholar]
- Benoist C., O'Hare K., Breathnach R., Chambon P. The ovalbumin gene-sequence of putative control regions. Nucleic Acids Res. 1980 Jan 11;8(1):127–142. doi: 10.1093/nar/8.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
- Burke R. L., Tekamp-Olson P., Najarian R. The isolation, characterization, and sequence of the pyruvate kinase gene of Saccharomyces cerevisiae. J Biol Chem. 1983 Feb 25;258(4):2193–2201. [PubMed] [Google Scholar]
- Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
- Curtis S. J., Epstein W. Phosphorylation of D-glucose in Escherichia coli mutants defective in glucosephosphotransferase, mannosephosphotransferase, and glucokinase. J Bacteriol. 1975 Jun;122(3):1189–1199. doi: 10.1128/jb.122.3.1189-1199.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dobson M. J., Tuite M. F., Roberts N. A., Kingsman A. J., Kingsman S. M., Perkins R. E., Conroy S. C., Fothergill L. A. Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae. Nucleic Acids Res. 1982 Apr 24;10(8):2625–2637. doi: 10.1093/nar/10.8.2625. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Entian K. D., Kopetzki E., Fröhlich K. U., Mecke D. Cloning of hexokinase isoenzyme PI from Saccharomyces cerevisiae: PI transformants confirm the unique role of hexokinase isoenzyme PII for glucose repression in yeasts. Mol Gen Genet. 1984;198(2):50–54. doi: 10.1007/BF00328699. [DOI] [PubMed] [Google Scholar]
- Falco S. C., Botstein D. A rapid chromosome-mapping method for cloned fragments of yeast DNA. Genetics. 1983 Dec;105(4):857–872. doi: 10.1093/genetics/105.4.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Faye G., Leung D. W., Tatchell K., Hall B. D., Smith M. Deletion mapping of sequences essential for in vivo transcription of the iso-1-cytochrome c gene. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2258–2262. doi: 10.1073/pnas.78.4.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fletterick R. J., Bates D. J., Steitz T. A. The structure of a yeast hexokinase monomer and its complexes with substrates at 2.7-A resolution. Proc Natl Acad Sci U S A. 1975 Jan;72(1):38–42. doi: 10.1073/pnas.72.1.38. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gancedo J. M., Clifton D., Fraenkel D. G. Yeast hexokinase mutants. J Biol Chem. 1977 Jul 10;252(13):4443–4444. [PubMed] [Google Scholar]
- Guarente L., Lalonde B., Gifford P., Alani E. Distinctly regulated tandem upstream activation sites mediate catabolite repression of the CYC1 gene of S. cerevisiae. Cell. 1984 Feb;36(2):503–511. doi: 10.1016/0092-8674(84)90243-5. [DOI] [PubMed] [Google Scholar]
- Guarente L. Yeast promoters: positive and negative elements. Cell. 1984 Apr;36(4):799–800. doi: 10.1016/0092-8674(84)90028-x. [DOI] [PubMed] [Google Scholar]
- Henikoff S., Cohen E. H. Sequences responsible for transcription termination on a gene segment in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Aug;4(8):1515–1520. doi: 10.1128/mcb.4.8.1515. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holland J. P., Holland M. J. Structural comparison of two nontandemly repeated yeast glyceraldehyde-3-phosphate dehydrogenase genes. J Biol Chem. 1980 Mar 25;255(6):2596–2605. [PubMed] [Google Scholar]
- Holland J. P., Holland M. J. The primary structure of a glyceraldehyde-3-phosphate dehydrogenase gene from Saccharomyces cerevisiae. J Biol Chem. 1979 Oct 10;254(19):9839–9845. [PubMed] [Google Scholar]
- Lobo Z., Maitra P. K. Genetics of yeast hexokinase. Genetics. 1977 Aug;86(4):727–744. doi: 10.1093/genetics/86.4.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Messing J., Gronenborn B., Müller-Hill B., Hans Hopschneider P. Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3642–3646. doi: 10.1073/pnas.74.9.3642. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Montgomery D. L., Leung D. W., Smith M., Shalit P., Faye G., Hall B. D. Isolation and sequence of the gene for iso-2-cytochrome c in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jan;77(1):541–545. doi: 10.1073/pnas.77.1.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Platt T. Termination of transcription and its regulation in the tryptophan operon of E. coli. Cell. 1981 Apr;24(1):10–23. doi: 10.1016/0092-8674(81)90496-7. [DOI] [PubMed] [Google Scholar]
- Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
- Shoham M., Steitz T. A. The 6-hydroxymethyl group of a hexose is essential for the substrate-induced closure of the cleft in hexokinase. Biochim Biophys Acta. 1982 Aug 10;705(3):380–384. doi: 10.1016/0167-4838(82)90260-6. [DOI] [PubMed] [Google Scholar]
- Steitz T. A., Fletterick R. J., Anderson W. F., Anderson C. M. High resolution x-ray structure of yeast hexokinase, an allosteric protein exhibiting a non-symmetric arrangement of subunits. J Mol Biol. 1976 Jun 14;104(1):197–122. doi: 10.1016/0022-2836(76)90009-7. [DOI] [PubMed] [Google Scholar]
- Tomita M., Furthmayr H., Marchesi V. T. Primary structure of human erythrocyte glycophorin A. Isolation and characterization of peptides and complete amino acid sequence. Biochemistry. 1978 Oct 31;17(22):4756–4770. doi: 10.1021/bi00615a025. [DOI] [PubMed] [Google Scholar]
- Zaret K. S., Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. doi: 10.1016/0092-8674(82)90211-2. [DOI] [PubMed] [Google Scholar]
- Zaret K. S., Sherman F. Mutationally altered 3' ends of yeast CYC1 mRNA affect transcript stability and translational efficiency. J Mol Biol. 1984 Jul 25;177(1):107–135. doi: 10.1016/0022-2836(84)90060-3. [DOI] [PubMed] [Google Scholar]