Brain and muscle creatine kinase genes contain common TA-rich recognition protein-binding regulatory elements.
We have previously reported that the rat brain creatine kinase (ckb) gene promoter contains an AT-rich sequence that is a binding site for a protein called TARP (TA-rich recognition protein). This AT-rich segment is a positively acting regulatory element for the ckb promoter. A similar AT-rich DNA segment is found at the 3' end of the 5' muscle-specific enhancer of the rat muscle creatine kinase (ckm) gene and has been shown to be necessary for full muscle-specific enhancer activity. In this report, we show that TARP binds not only to the ckb promoter but also to the AT-rich segment at the 3' end of the muscle-specific ckm enhancer. A second, weaker TARP-binding site was identified in the ckm enhancer and lies at the 5' end of the minimal enhancer segment. TARP was found in both muscle cells (C2 and L6 myotubes) and nonmuscle (HeLa) cells and appeared to be indistinguishable from both sources, as judged by gel retardation and footprinting assays. The TARP-binding sites in the ckm enhancer and the ckb promoter were found to be functionally interchangeable. We propose that TARP is active in both muscle and nonmuscle cells and that it is one of many potential activators that may interact with muscle-specific regulators to determine the myogenic phenotype.
Horlick
R A
RA
E. I. du Pont de Nemours and Co., Inc., Wilmington, Delaware 19880-0328.
Hobson
G M
GM
Patterson
J H
JH
Mitchell
M T
MT
Benfield
P A
PA
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D003402:Creatine Kinase / Q000235:genetics*; D003850:Deoxyribonuclease I; D005796:Genes; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology*; D009710:Nucleotide Mapping; D015345:Oligonucleotide Probes; D011401:Promoter Regions, Genetic; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014162:Transfection
D015345:Oligonucleotide Probes; D003402:Creatine Kinase; D003850:Deoxyribonuclease I