Positive and negative regulatory elements of the rabbit embryonic εglobin gene revealed by an improved multiple alignment program and functional analysis
Journal
Mitochondrial DNA
Journal Volume
4
Journal Issue
3
Pages
163-176
Date Issued
1993
Author(s)
Abstract
The εglobin genes of mammals are expressed in early embryos, but are silenced during fetal and adult erythropoiesis. As a guide to defining the regulatory elements involved in this developmental switch, we have searched the sequences of εglobin genes from different mammals for highly conserved segments. The search was facilitated by the development of a new program, called yama, to generate a multiple alignment of very long sequences using an improved scoring scheme. This allowed us to generate a multiple alignment of sequences from a more divergent group than previously analyzed, as demonstrated here for representatives of four mammalian orders. In parallel experiments, we constructed a series of deletion mutations in the 5' flank of the rabbit εglobin gene and tested their effect on an εglobin-luciferase hybrid reporter gene. These results show that 121 bp of 5' flank, containing CACC, CCAAT and ATA motifs, is sufficient for expression in erythroid K562 cells. Both positive and negative cis-acting control sequences are located between 218 and 394 bp 5' to the cap site, in a region previously proposed to be a silencer. The positive regulatory sequence contains conserved binding sites for the nuclear protein YY1 adjacent to another highly conserved sequence. The negative element contains a conserved sequence followed by a purine-rich segment. This analysis maps the upstream control sequences more precisely and points to a very complex regulatory scheme for this gene. © 1993 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.
Subjects
E-globin; Gene regulation; Luciferase; Multiple sequence alignment; Negative regulation; Positive; RNA analyses
Type
journal article