PROTEIN-DNA INTERACTIONS AT PUTATIVE REGULATORY REGIONS OF 2 COORDINATELY EXPRESSED GENES, MSP130 AND PM27, DURING SKELETOGENESIS IN SEA-URCHIN EMBRYOS
V. Raman et al., PROTEIN-DNA INTERACTIONS AT PUTATIVE REGULATORY REGIONS OF 2 COORDINATELY EXPRESSED GENES, MSP130 AND PM27, DURING SKELETOGENESIS IN SEA-URCHIN EMBRYOS, The International journal of developmental biology, 37(4), 1993, pp. 499-507
Development of the primary mesenchyme cells (PMCs) of the sea urchin e
mbryo, which give rise to the larval skeleton, involves the coordinate
onset of expression of several structural genes. As part of an effort
to identify cis-acting elements that might play a role in this regula
tory event, co-regulated genes were examined by two approaches. First,
they were compared for conserved sequence elements. Four conserved el
ements were found as a cluster in all three genes examined, suggesting
a regulatory role. Second, as a test for potential function, the puta
tive regulatory regions of two of these genes were examined for protei
n binding sites. DNase I protection and gel mobility shift assays were
used to: 1) identify several nuclear protein binding sites in these r
egions, two of which correspond to conserved elements among the genes;
2) demonstrate that the developmental time of appearance of the prote
ins that interact with these sites corresponds to the time of activati
on of the genes; and 3) show that two of the conserved sequence elemen
ts shared by these genes compete for the same binding proteins. These
data identify putative regulatory elements, whose specific roles in th
e coordinate regulation of PMC-expressed genes can now be addressed di
rectly using appropriate transgenic expression constructs.