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Molecular and Cellular Biology, August 1999, p. 5557-5564, Vol. 19, No. 8
Department of Molecular and Cell Biology,
University of California at Berkeley, Berkeley, California 94720
Received 23 February 1999/Returned for modification 7 April
1999/Accepted 4 May 1999
The initiation of translation in eukaryotes requires several
multisubunit complexes, including eukaryotic translation initiation factor 4F (eIF4F). In higher eukaryotes eIF4F is composed of the cap
binding protein eIF4E, the adapter protein eIF4G, and the RNA-stimulated ATPase eIF4A. The association of eIF4A with
Saccharomyces cerevisiae eIF4F has not yet been
demonstrated, and therefore the degree to which eIF4A's conserved
function relies upon this association has remained unclear. Here we
report an interaction between yeast eIF4G and eIF4A. Specifically, we
found that the growth arrest phenotype associated with three
temperature-sensitive alleles of yeast eIF4G2 was suppressed by excess
eIF4A and that this suppression was allele specific. In addition, in
vitro translation extracts derived from an eIF4G2 mutant strain could
be heat inactivated, and this inactivation could be reversed upon the
addition of recombinant eIF4A. Finally, in vitro binding between yeast
eIF4G and eIF4A was demonstrated, as was diminished binding between
mutant eIF4G2 proteins and eIF4A. In total, these data indicate that
yeast eIF4G and eIF4A physically associate and that this association
performs an essential function.
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Eukaryotic Translation Initiation Factors 4G and 4A
from Saccharomyces cerevisiae Interact Physically and
Functionally
*
Corresponding author. Mailing address: Department of
Molecular and Cell Biology, 401 Barker Hall, University of California at Berkeley, Berkeley, CA 94720. Phone: (510) 643-7698. Fax: (510) 643-5035. E-mail: asachs{at}uclink4.berkeley.edu.
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