Structure, sequence and expression of the mouse Cx43 gene encoding connexin 43.
Gap junctions, membrane channels that mediate the diffusion of ions and small molecules between cells, are hypothesized to play a role in development and growth regulation. The Cx43 gene (encoding connexin 43) is one member of the gap junction gene family whose transcripts are expressed in a highly regionalized manner during mouse development. We cloned and sequenced Cx43 cDNAs from a 7.5-day mouse embryo cDNA library. These cDNA clones encode the authentic 43-kDa connexin. Analysis of RNA isolated from different regions of the 7.5-day mouse embryo revealed that Cx43 transcripts are differentially expressed, with expression detected in the embryo proper, but not in the extraembryonic region containing the ectoplacental cone. Using one of the newly isolated mouse Cx43 cDNA probes, we screened a mouse genomic DNA library and cloned the Cx43 gene. Restriction mapping and sequencing of the cloned genomic inserts revealed that Cx43 contains two exons and a 10.5-kb intron located in the 5' untranslated region (5'-UTR). We mapped the Cx43 transcription start point (tsp) by RNase protection and primer extension analyses and showed that transcripts expressed in the 7.5-day mouse embryo and in adult tissues are initiated from the same tsp. The DNA sequence immediately upstream from the tsp contains a putative AP1-binding site and a degenerate TATA consensus sequence. A comparison of mouse, rat, human and bovine Cx43s showed that the 3'-UTR has an unexpectedly high degree of sequence homology. This includes conservation of four AUUUA motifs, a sequence associated with transcript instability in immediate early genes.(ABSTRACT TRUNCATED AT 250 WORDS)
Biology Department, University of Pennsylvania, Philadelphia 19104-6017.
R
R
Sullivan
C
C
Ruangvoravat
D
D
Joo
J
J
Morgan
B L
BL
Wang
X K
XK
Wang
C W
CW
Lo
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017630:Connexins; D004247:DNA; D015870:Gene Expression; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D014158:Transcription, Genetic
D017630:Connexins; D008565:Membrane Proteins; D004247:DNA