Roles of Ebp2 and ribosomal protein L36 in ribosome biogenesis in Saccharomyces cerevisiae
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Ebp2 plays an essential role in biogenesis of 60S ribosomal subunits. We determined the genetic interactions between EBP2 and RPL36A/B, which encodes ribosomal protein L36a/b. RPL36A/B was a multicopy suppressor to ebp2 mutants, and the suppression was not common to defects in ribosome biogenesis resulting from other mutations of assembly factors. Disruption of RPL36A or RPL36B caused synthetic enhancement of the growth defect of the ebp2-14 allele at high temperatures. Disruption of RPL36B led to a more severe growth defect than that of RPL36A due to imbalances in the expression levels of the duplicated genes. Primer-extension analysis revealed that L36a/b is required for the processing of 27SA2, 27SA3, and 27SBL pre-rRNAs. Two-hybrid analysis indicated that Ebp2 interacts with ribosomal proteins L36a/b, L34a/b, and L8, which in mature ribosomes are located adjacent to each other in close proximity to the 3′ end of 5.8S rRNA. These results suggest that Ebp2 functions cooperatively with ribosomal proteins L36, L34, and L8 in biogenesis of the 60S ribosomal subunit.
KeywordsRibosome biogenesis EBP2 Ribosomal protein L36a/b Ribosomal protein L34a/b Duplicated genes
lexA binding domain
Gal4 activation domain
We thank K. Kanda for screening of multicopy suppressors, Dr. E. Tsuchiya for the yeast genomic library, Dr. J. L. Woolford, Jr., Dr. D. Shore, and Dr. K. Tanaka for plasmids, and Dr. K. Funato for helpful discussion. This work was supported by JSPS KAKENHI Grant Number 25450516 (to K.M.).
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