Abstract | BACKGROUND: RESULTS: To suppress eIF2B mutations, S. cerevisiae genomic DNA library was constructed in high-copy vector (YEp24) and transformed into eIF2B mutant S. cerevisiae strains. The library was screened for wild-type genes rescuing S. cerevisiae (Slg-) and (Gcd-) phenotypes. A genomic clone, Suppressor-I (Sup-I), rescued S. cerevisiae Slg- and Gcd- phenotypes (gcd7-201 gcn2∆). The YEp24/Sup-I construct contained truncated TAN1, full length EMC4, full length YGL230C, and truncated SAP4 genes. Full length EMC4 (chaperone protein) gene was sub-cloned into pEG (KG) yeast expression vector and overexpressed in gcd7-201 gcn2∆ strain which suppressed the Slg- and Gcd- phenotype. A GST-Emc4 fusion protein of 47 kDa was detected by western blotting using α-GST antibodies. Suppression was specific to gcd7-201 gcn2∆ mutation in eIF2Bβ and Gcd1-502 gcn2∆ in eIF2Bγ subunit. Emc4p overexpression also protected the wild type and mutant (gcd7-201 gcn2∆, GCD7 gcn2∆, and GCD7 GCN2∆) strains from H2O2, ethanol, and caffeine stress. CONCLUSIONS: Our results suggest that Emc4p is involved in eIF2B-mediated translational regulation under stress and could provide an amenable tool to understand the eIF2B-mediated defects.
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Authors | Sonum Sharma, Anuradha Sourirajan, David J Baumler, Kamal Dev |
Journal | Journal, genetic engineering & biotechnology
(J Genet Eng Biotechnol)
Vol. 18
Issue 1
Pg. 15
(Jun 01 2020)
ISSN: 2090-5920 [Electronic] Netherlands |
PMID | 32476094
(Publication Type: Journal Article)
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