Functional characterization of the promoter region of the chicken elongation factor-2 gene.
Elongation factor 2 (EF-2) plays a key role in the essential process of protein synthesis by translocating tRNAs from the ribosomal A- and P-sites to the P- and E-sites. EF-2 regulates the outcome of protein synthesis in mammalian cells. This report demonstrates that chicken EF-2 protein levels are dependent on transcription in 8-bromo-cAMP, insulin and phorbol ester-treated cells. In order to delineate functional domains that control chicken EF-2 gene transcription, the 5'-flanking region of the chicken EF-2 promoter was analyzed. Deletion constructs from -550 and -86 had the same basal level promoter activity as the whole EF-2 promoter. The sequence between nucleotides -700 and -550 was determined to be a regulatory region for the chicken EF-2 basal promoter activity. The region between -700 and -550 has a negative regulatory region and two regulatory proteins (I, II). 8-bromo-cAMP increased chicken EF-2 promoter activity (-700/+102) in Rat 1 HIR fibroblast cells more than insulin and phorbol ester treatment. Binding of protein I and II were decreased by 8-bromo-cAMP but restored by a protein kinase A inhibitor (KT5720). GATA consensus sequence oligonucleotide and fragment -86/-50 prevented protein II binding of fragment -700/-550. This result suggested that protein II is a GATA-like protein. These observations provide a novel regulatory mechanism for the EF-2 promoter.
Lim
Eun Jin
EJ
Molecular and Cell Nutrition Laboratory, Department of Animal and Food Science, University of Kentucky, Lexington, Kentucky 40536, USA. eunjinlim@uky.edu
Kim
Choong Won
CW
D020121:5' Untranslated Regions / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D002645:Chickens / Q000235:genetics*; D050982:GATA1 Transcription Factor / Q000502:physiology; D008969:Molecular Sequence Data; D020652:Peptide Elongation Factor 2 / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000502:physiology*; D051381:Rats
D020121:5' Untranslated Regions; D050982:GATA1 Transcription Factor; D020652:Peptide Elongation Factor 2