Naturally occurring antisense transcripts are present in chick embryo chondrocytes simultaneously with the down-regulation of the alpha 1 (I) collagen gene.
It has previously been shown that very low steady state levels of alpha 1 (I) collagen mRNA are present in chick embryo sternal chondrocytes (Askew, G. R., Wang, S., and Lukens, L. N. (1991) J. Biol. Chem. 266, 16834-16841), yet nuclear run-on experiments with double-stranded cDNA probes indicated a high transcription rate at this locus. These findings were investigated in this study using single-stranded probes, where nuclear run-on experiments showed that antisense transcription of the alpha 1 (I) collagen gene was occurring in chondrocytes, while sense strand transcription was down-regulated. Treatment of these chondrocytes with 5-bromo-2'-deoxyuridine (BrdU), which causes the cells to resemble their mesenchymal precursors, resulted in an antiparallel situation, where antisense transcription was lost, and instead, sense strand transcription was acquired, suggesting that the reverse switch from sense to antisense transcription occurs during chondrogenesis. Very large (> 10 kilobases) and heterogeneous antisense transcripts of moderate stability were shown to span both ends of the gene in chondrocytes, while their absence was shown in BrdU-treated chondrocytes, chick embryo fibroblasts, and a variety of other tissues. The function of these antisense transcripts is so far unknown, but their unusual chondrocyte-specific appearance, concurrent with little or no sense strand transcription, suggests a possible functional role in the down-regulation of the alpha 1 (I) collagen gene.
Molecular Biology and Biochemistry Department, Wesleyan University, Middletown, Connecticut 06459-0175.
C M
CM
Farrell
L N
LN
Lukens
D000818:Animals; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002356:Cartilage / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002642:Chick Embryo; D003001:Cloning, Molecular; D003094:Collagen / Q000096:biosynthesis* / Q000235:genetics; D015342:DNA Probes; D018076:DNA, Complementary; D005786:Gene Expression Regulation; D008970:Molecular Weight; D010957:Plasmids; D016372:RNA, Antisense / Q000096:biosynthesis*; D015183:Restriction Mapping; D012886:Skull / Q000378:metabolism; D014158:Transcription, Genetic
D015342:DNA Probes; D018076:DNA, Complementary; D016372:RNA, Antisense; D003094:Collagen; D001973:Bromodeoxyuridine