Abstract
We have identified a yeast protein that resembles actins from other eucaryotes in its tight binding to pancreatic deoxyribonuclease I, its copolymerizaton with purified muscle actin, its one-dimensional peptide map, and its apparent polymerization into 7-nm filaments. The yeast actin-like protein yielded a single spot on two-dimensional polyacrylamide gel electrophoresis, suggesting that a single protein species was present. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the actin-like protein had an apparent molecular weight of 45,000 compared with 42,000 for muscle actin. In an attempt to identify the messenger ribonucleic acid coding for the actin-like protein, yeast polyadenylic acid-rich ribonucleic acid was translated in wheat germ and reticulocyte cell-free protein-synthesizing systems. The actin-like protein was identified among the translation products of the reticulocyte system by its tight binding to deoxyribonuclease I, its comigration with the in vivo-synthesized actin-like protein during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, an the similarity of its peptide map to that of the in vivo-synthesized protein. A yeast protein synthesized in the wheat-germ system was also found to bind to deoxyribonuclease I and to copolymerize with muscle actin. However, its apparent molecular weight was about 35,000, suggesting that it was a product either of incomplete translation or of proteolytic cleavage of the actin-like protein.
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Selected References
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- Adams J., Hansche P. E. Population studies in microorganisms. I. Evolution of diploidy in Saccharomyces cerevisiae. Genetics. 1974 Feb;76(2):327–338. doi: 10.1093/genetics/76.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Allen E. D., Sussman A. S. Presence of an actin-like protein in mycelium of Neurospora crassa. J Bacteriol. 1978 Aug;135(2):713–716. doi: 10.1128/jb.135.2.713-716.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Benveniste K., Wilczek J., Ruggieri A., Stern R. Translation of collagen messenger RNA in a cell-free system derived from wheat germ. Biochemistry. 1976 Feb 24;15(4):830–835. doi: 10.1021/bi00649a016. [DOI] [PubMed] [Google Scholar]
- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Byers B., Goetsch L. A highly ordered ring of membrane-associated filaments in budding yeast. J Cell Biol. 1976 Jun;69(3):717–721. doi: 10.1083/jcb.69.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clarke M., Spudich J. A. Nonmuscle contractile proteins: the role of actin and myosin in cell motility and shape determination. Annu Rev Biochem. 1977;46:797–822. doi: 10.1146/annurev.bi.46.070177.004053. [DOI] [PubMed] [Google Scholar]
- Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
- Furthmayr H., Timpl R. Characterization of collagen peptides by sodium dodecylsulfate-polyacrylamide electrophoresis. Anal Biochem. 1971 Jun;41(2):510–516. doi: 10.1016/0003-2697(71)90173-4. [DOI] [PubMed] [Google Scholar]
- Gallis B. M., McDonnell J. P., Hopper J. E., Young E. T. Translation of poly(riboadenylic acid)-enriched messenger RNAs from the yeast, Saccharomyces cerevisiae, in heterologous cell-free systems. Biochemistry. 1975 Mar 11;14(5):1038–1046. doi: 10.1021/bi00676a024. [DOI] [PubMed] [Google Scholar]
- Garrels J. I., Gibson W. Identification and characterization of multiple forms of actin. Cell. 1976 Dec;9(4 Pt 2):793–805. doi: 10.1016/0092-8674(76)90142-2. [DOI] [PubMed] [Google Scholar]
- Gozes I., Schmitt H., Littauer U. Z. Translation in vitro of rat brain messenger RNA coding for tubulin and actin. Proc Natl Acad Sci U S A. 1975 Feb;72(2):701–705. doi: 10.1073/pnas.72.2.701. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hartwell L. H. Cell division from a genetic perspective. J Cell Biol. 1978 Jun;77(3):627–637. doi: 10.1083/jcb.77.3.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hartwell L. H., Mortimer R. K., Culotti J., Culotti M. Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants. Genetics. 1973 Jun;74(2):267–286. doi: 10.1093/genetics/74.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hartwell L. H. Saccharomyces cerevisiae cell cycle. Bacteriol Rev. 1974 Jun;38(2):164–198. doi: 10.1128/br.38.2.164-198.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holland M. J., Holland J. P. Isolation and identification of yeast messenger ribonucleic acids coding for enolase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase. Biochemistry. 1978 Nov 14;17(23):4900–4907. doi: 10.1021/bi00616a007. [DOI] [PubMed] [Google Scholar]
- Hunter T., Garrels J. I. Characterization of the mRNAs for alpha-, beta- and gamma-actin. Cell. 1977 Nov;12(3):767–781. doi: 10.1016/0092-8674(77)90276-8. [DOI] [PubMed] [Google Scholar]
- Koteliansky V. E., Glukhova M. A., Bejanian M. V., Surguchov A. P., Smirnov V. N. Isolation and characterization of actin-like protein from yeast Saccharomyces cerevisiae. FEBS Lett. 1979 Jun 1;102(1):55–58. doi: 10.1016/0014-5793(79)80927-8. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lazarides E., Lindberg U. Actin is the naturally occurring inhibitor of deoxyribonuclease I. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4742–4746. doi: 10.1073/pnas.71.12.4742. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LeStourgeon W. M., Rusch H. P. Localization of nucleolar and chromatin residual acidic protein changes during differentiation in Physarum polycephalum. Arch Biochem Biophys. 1973 Mar;155(1):144–158. doi: 10.1016/s0003-9861(73)80017-7. [DOI] [PubMed] [Google Scholar]
- Luduena R. F., Woodward D. O. Alpha- and beta-tubulin: separation and partial sequence analysis. Ann N Y Acad Sci. 1975 Jun 30;253:272–283. doi: 10.1111/j.1749-6632.1975.tb19206.x. [DOI] [PubMed] [Google Scholar]
- Mortimer R. K., Hawthorne D. C. Genetic mapping in yeast. Methods Cell Biol. 1975;11:221–233. doi: 10.1016/s0091-679x(08)60325-8. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
- Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
- Pringle J. R. Methods for avoiding proteolytic artefacts in studies of enzymes and other proteins from yeasts. Methods Cell Biol. 1975;12:149–184. doi: 10.1016/s0091-679x(08)60956-5. [DOI] [PubMed] [Google Scholar]
- Roberts B. E., Paterson B. M. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330–2334. doi: 10.1073/pnas.70.8.2330. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger J. W. Presence of actin during chromosomal movement. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2451–2455. doi: 10.1073/pnas.72.6.2451. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
- Storti R. V., Rich A. Chick cytoplasmic actin and muscle actin have different structural genes. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2346–2350. doi: 10.1073/pnas.73.7.2346. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tse T. P., Taylor J. M. Translation of albumin messenger RNA in a cell-free protein-synthesizing system derived from wheat germ. J Biol Chem. 1977 Feb 25;252(4):1272–1278. [PubMed] [Google Scholar]
- Tung J. S., Knight C. A. Effect of charge on the determination of molecular weight of proteins by gel electrophoresis in SDS. Biochem Biophys Res Commun. 1971 Mar 19;42(6):1117–1121. doi: 10.1016/0006-291x(71)90020-9. [DOI] [PubMed] [Google Scholar]
- Vandekerckhove J., Weber K. The amino acid sequence of Physarum actin. Nature. 1978 Dec 14;276(5689):720–721. doi: 10.1038/276720a0. [DOI] [PubMed] [Google Scholar]
- Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]
- Wilkinson L. E., Pringle J. R. Transient G1 arrest of S. cerevisiae cells of mating type alpha by a factor produced by cells of mating type a. Exp Cell Res. 1974 Nov;89(1):175–187. doi: 10.1016/0014-4827(74)90200-6. [DOI] [PubMed] [Google Scholar]