Abstract
A protein-synthesizing system consisting of ribosomes and supernatant of undeveloped Artemia saline embryos is used for assay of mRNA translation and initiation factors. This system contains the components needed for chain elongation but has low levels of mRNA and initiation factors. Exogenous mRNA is readily translated upon addition of high-speed supernatant or ribosomal salt wash of developing embryos as a source of initiation factors (IF). This requirement can be largely satisfied by a mixture of the reticulocyte factors IF-MP, IF-M3, IF-M2A, and IF-M2B. A. salina IF-M1, which is present in undeveloped embryo supernatant, can be inactivated by A. salina IF-M1 antibody without affecting translation.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bonanou-Tzedaki S. A., Pragnell I. B., Arnstein H. R. Presence of haemoglobin messenger RNA in the postribosomal supernatant of rabbit reticulocytes and conditions necessary for its translation. FEBS Lett. 1972 Oct 1;26(1):77–82. doi: 10.1016/0014-5793(72)80546-5. [DOI] [PubMed] [Google Scholar]
- Cashion L. M., Stanley W. M., Jr Two eukaryotic initiation factors (IF-I and IF-II) of protein synthesis that are required to form an initiation complex with rabbit reticulocyte ribosomes. Proc Natl Acad Sci U S A. 1974 Feb;71(2):436–440. doi: 10.1073/pnas.71.2.436. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Caskey T., Leder P., Moldave K., Schlessinger D. Translation: its mechanism and control. Science. 1972 Apr 14;176(4031):195–197. doi: 10.1126/science.176.4031.195. [DOI] [PubMed] [Google Scholar]
- Filipowicz W., Sierra J. M., Ochoa S. Polypeptide chain initiation in eukaryotes: initiation factor MP in Artemia salina embryos. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3947–3951. doi: 10.1073/pnas.72.10.3947. [DOI] [PMC free article] [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]
- Levin D. H., Kyner D., Acs G. Protein synthesis initiation in eukaryotes. Characterization of ribosomal factors from mouse fibroblasts. J Biol Chem. 1973 Sep 25;248(18):6416–6425. [PubMed] [Google Scholar]
- Merrick W. C., Kemper W. M., Anderson W. F. Purification and characterization of homogeneous initiation factor M2A from rabbit reticulocytes. J Biol Chem. 1975 Jul 25;250(14):5556–5562. [PubMed] [Google Scholar]
- Merrick W., Graf H., Anderson W. F. Preparation of protein synthesis initiation factors IF-M1, IF-M2A, and IF-M2B from rabbit reticulocytes. Methods Enzymol. 1974;30:128–136. doi: 10.1016/0076-6879(74)30015-8. [DOI] [PubMed] [Google Scholar]
- Nombela C., Nombela N. A., Ochoa S., Merrick W. C., Anderson F. Nature of eukaryotic proteins required for joining of 40S and 60S ribosomal subunits. Biochem Biophys Res Commun. 1975 Mar 17;63(2):409–416. doi: 10.1016/0006-291x(75)90703-2. [DOI] [PubMed] [Google Scholar]
- Nombela C., Ochoa S. Conformational control of the interaction of eukaryotic elongation factors EF-1 and EF-2 with ribosomes. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3556–3560. doi: 10.1073/pnas.70.12.3556. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PORTER R. R. Separation and isolation of fractions of rabbit gamma-globulin containing the antibody and antigenic combining sites. Nature. 1958 Sep 6;182(4636):670–671. doi: 10.1038/182670a0. [DOI] [PubMed] [Google Scholar]
- Prichard P. M., Anderson W. F. Preparation of rabbit reticulocyte initiation factor IF-M3. Methods Enzymol. 1974;30:136–141. doi: 10.1016/0076-6879(74)30016-x. [DOI] [PubMed] [Google Scholar]
- Prichard P. M., Gilbert J. M., Shafritz D. A., Anderson W. F. Factors for the initiation of haemoglobin synthesis by rabbit reticulocyte ribosomes. Nature. 1970 May 9;226(5245):511–514. doi: 10.1038/226511a0. [DOI] [PubMed] [Google Scholar]
- Schreier M. H., Staehelin T. Initiation of eukaryotic protein synthesis: (Met-tRNA f -40S ribosome) initiation complex catalysed by purified initiation factors in the absence of mRNA. Nat New Biol. 1973 Mar 14;242(115):35–38. doi: 10.1038/newbio242035a0. [DOI] [PubMed] [Google Scholar]
- Shafritz D. A., Prichard P. M., Gilbert J. M., Merrick W. C., Anderson W. F. Separation of reticulocyte initiation factor M 2 activity into two components. Proc Natl Acad Sci U S A. 1972 Apr;69(4):983–987. doi: 10.1073/pnas.69.4.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sierra J. M., Meier D., Ochoa S. Effect of development on the translation of messenger RNA in Artemia salina embryos. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2693–2697. doi: 10.1073/pnas.71.7.2693. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strycharz W. A., Ranki M., Dahl H. H. A high-molecular-weight protein component required for natural messenger translation in ascites tumor cells. Eur J Biochem. 1974 Oct 1;48(1):303–310. doi: 10.1111/j.1432-1033.1974.tb03769.x. [DOI] [PubMed] [Google Scholar]
- Wigle D. T., Smith A. E. Specificity in initiation of protein synthesis in a fractionated mammalian cell-free system. Nat New Biol. 1973 Apr 4;242(118):136–140. doi: 10.1038/newbio242136a0. [DOI] [PubMed] [Google Scholar]
- Zasloff M., Ochoa S. A supernatant factor involved in initiation complex formation with eukaryotic ribosomes. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3059–3063. doi: 10.1073/pnas.68.12.3059. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zasloff M., Ochoa S. Polypeptide chain initiation in eukaryotes. IV. Purification and properties of supernatant initiation factor from Artemia salina embryos. J Mol Biol. 1973 Jan;73(1):65–76. doi: 10.1016/0022-2836(73)90159-9. [DOI] [PubMed] [Google Scholar]