DOI: 10.1111/j.1432-1033.1989.tb14793.x. A novel role for aminoacyl‐tRNA synthetases in the regulation of polypeptide chain initiation

A novel role for aminoacyl‐tRNA synthetases in the regulation of polypeptide chain initiation

10.1111/j.1432-1033.1989.tb14793.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

Exposure of the temperature‐sensitive leucyl‐tRNA synthetase mutant of Chinese hamster ovary cells, tsH1, to the non‐permissive temperature of 39.5°C results in a rapid inhibition of polypeptide chain initiation. This inhibition is caused by a reduced ability of the eukaryotic initiation factor eIF‐2 to participate in the formation of eIF‐2 · GTP · Met‐tRNAf ternary complexes and thus in the formation of 43S ribosomal pre‐initiation complexes. Associated with this decreased eIF‐2 activity is an increased phosphorylation of the eIF‐2α subunit. It has previously been shown in other systems that phosphorylation of eIF‐2α slows the rate of recycling of eIF‐2 · GDP to eIF‐2 · GTP catalysed by the guanine nucleotide exchange factor eIF‐2B. We show here that phosphorylation of eIF‐2α by the reticulocyte haem‐controlled repressor also inhibits eIF‐2B activity in cell‐free extracts derived from tsH1 cells. Thus the observed increased phosphorylation of eIF‐2α at the non‐permissive temperature in this system is consistent with impaired recycling of eIF‐2 in vivo.Using a single‐step temperature revertant of tsH1 cells, TR‐3 (which has normal leucyl‐tRNA synthetase activity at 39.5°C), we demonstrate here that all inhibition of eIF‐2 function reverts together with the synthetase mutation. This establishes the close link between synthetase function and eIF‐2 activity. In contrast, recharging tRNALeuin vivo in tsH1 cells at 39.5°C by treatment with a low concentration of cycloheximide failed to reverse the inhibition of eIF‐2 function. This indicates that tRNA charging per se is not involved in the regulatory mechanism.Our data indicate a novel role for aminoacyl‐tRNA synthetases in the regulation of eIF‐2 function mediated through phosphorylation of the α subunit of this factor. However, in spite of the fact that cell‐free extracts from Chinese hamster ovary cells contain protein kinase and phosphatase activities active against either exogenous or endogenous eIF‐2α, we have been unable to show any activation of kinase or inactivation of phosphatase following incubation of the cells at 39.5°C.

Bibliography

POLLARD, J. W., GALPINE, A. R., & CLEMENS, M. J. (1989). A novel role for aminoacylâtRNA synthetases in the regulation of polypeptide chain initiation. European Journal of Biochemistry, 182(1), 1â9. Portico.

Authors 3

Jeffrey W. POLLARD (first)
Angela R. GALPINE (additional)
Michael J. CLEMENS (additional)

References 38 Referenced 22

{'key': 'e_1_2_3_2_2', 'first-page': '191', 'volume-title': 'Control of proliferation in animal cells', 'author': 'Stanners C. P.', 'year': '1974'} / Control of proliferation in animal cells by Stanners C. P. (1974)
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{'key': 'e_1_2_3_15_2', 'first-page': '47', 'volume': '4', 'author': 'Pollard J. W.', 'year': '1988', 'journal-title': 'Methods Mol. Biol.'} / Methods Mol. Biol. by Pollard J. W. (1988)
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{'key': 'e_1_2_3_28_2', 'first-page': '733', 'volume-title': 'Ribosomes', 'author': 'Cashel M.', 'year': '1974'} / Ribosomes by Cashel M. (1974)
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10.1016/S0021-9258(19)86057-X

Dates

Type	When
Created	20 years, 5 months ago (March 3, 2005, 10:09 p.m.)
Deposited	1 year, 9 months ago (Oct. 31, 2023, 8:59 a.m.)
Indexed	1 year, 9 months ago (Nov. 1, 2023, 3:21 a.m.)
Issued	36 years, 2 months ago (June 1, 1989)
Published	36 years, 2 months ago (June 1, 1989)
Published Online	20 years, 5 months ago (March 3, 2005)
Published Print	36 years, 2 months ago (June 1, 1989)

Funders 0

None

BibTeX

@article{POLLARD_1989, title={A novel role for aminoacyl‐tRNA synthetases in the regulation of polypeptide chain initiation}, volume={182}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1989.tb14793.x}, DOI={10.1111/j.1432-1033.1989.tb14793.x}, number={1}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={POLLARD, Jeffrey W. and GALPINE, Angela R. and CLEMENS, Michael J.}, year={1989}, month=jun, pages={1–9} }

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