Molecular Biology and Evolution, Vol 15, 312-325, Copyright © 1998 by Society for Molecular Biology and Evolution
N Singh, KW Barbour and FG Berger
Transcription of the murine D7Rp2e gene is highly variable among species of
the genus Mus, indicating that extensive modifications in the gene's
regulatory elements have occurred during evolution. Since promoter regions
are well known to harbor cis-acting information that controls gene
transcription, we compared the sequence and function of the D7Rp2e promoter
in several Mus species, with the goal of understanding the molecular
mechanisms underlying the interspecies variations in expression. Three
overlapping binding sites for nuclear factors (sites A, B, and C in
proximal to distal order) were identified about 300 bp upstream of the
transcriptional start site. The sequences of these sites differ between the
species Mus domesticus and M. pahari, which exhibit distinct D7Rp2e
expression phenotypes. Site A binds a factor called RPBF-I; sites B and C
bind a distinct factor that is termed RPBF-II and is likely a member of the
NF-I family of transcription factors. DNase I footprinting experiments with
the M. domesticus promoter show that binding of RPBF-II at site B is very
strong, while binding of RPBF-I and RPBF-II at sites A and C, respectively,
is weak; in contrast, with the M. pahari promoter, factor binding at sites
A and C is strong, while that at site B is weak. These differences in
patterns of binding-site occupancy derive from changes in the affinities of
individual sites for their cognate nuclear factors. Transient transfection
experiments indicate that the M. pahari binding pattern is capable of
repressing transcription of a linked reporter. Such repression may
contribute to the differences in D7Rp2e expression between the two species.
We suggest that the species- specific footprinting patterns represent the
existence of a dynamic equilibrium between two states of nuclear factor
binding, the nature of which can be modified during evolution to result in
new patterns of gene transcription.
ORIGINAL ARTICLE
Evolution of transcriptional regulatory elements within the promoter of a mammalian gene
Department of Biological Sciences, University of South Carolina, Columbia 29208, USA.
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