Characterization of an antisense Inr element in the eIF-2 alpha gene.

摘要: We recently discovered an opposing initiator promoter (Inr) downstream of the sense region eIF-2 alpha gene (Silverman, T., Noguchi, M., and Safer, B. (1992) J. Biol. Chem. 267, 9738-9742). By reverse transcriptase/polymerase chain reaction analysis G0 activated (G1) T-lymphocyte RNAs, overlapping antisense transcripts are now identified. Sense transcription proceeds from left to right generate alpha-mRNA; RNA, having a sequence complementary mRNA. Upstream indicates position 5' relative start site. Using DNase I footprint EMSA, we have found potential cis-regulatory immediately upstream Inr which binds 43-kDa protein. In addition conferring protection against (+457 +474), factor also generates hypersensitive sites directly over (+447 +457). Insertion into luciferase reporter construct increases expression 150-fold. While mutation conserved decreases activity by 50%, binding site inhibits 20%. orientation 80%. The Inr-associated protein may be involved in allowing access RNA polymerase II complexes ot initiation this TATA-less gene. A model for regulation involving rapid degradation dsRNA generated activities two promoters is proposed.