Positive and negative regulatory elements of the rabbit embryonic ϵ-globin gene revealed by an improved multiple alignment program and functional analysis
| Autor: | Doug Price, Nikola Stojanovic, Webb Miller, Tim Zeigler, Kun-Mao Chao, John Jackson, Michael Adamkiewicz, Ross C. Hardison |
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| Rok vydání: | 1993 |
| Předmět: |
Molecular Sequence Data
Biology Transfection Biochemistry Cell Line Embryonic and Fetal Development Endocrinology Genes Regulator Genetics Animals Humans Luciferase Molecular Biology Gene Conserved Sequence YY1 Transcription Factor Sequence Deletion Mammals Regulation of gene expression Reporter gene Binding Sites Multiple sequence alignment Base Sequence Functional analysis DNA Embryonic stem cell Globins DNA-Binding Proteins Gene Expression Regulation Erythroid-Specific DNA-Binding Factors Erythropoiesis Rabbits Sequence Alignment Software Transcription Factors |
| Zdroj: | DNA Sequence. 4:163-176 |
| ISSN: | 1042-5179 |
| DOI: | 10.3109/10425179309015629 |
| Popis: | The epsilon-globin 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 epsilon-globin 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 epsilon-globin gene and tested their effect on an epsilon-globin-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. |
| Databáze: | OpenAIRE |
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