PTRF (polymerase I and transcript-release factor) is tissue-specific and interacts with the BFCOL1 (binding factor of a type-I collagen promoter) zinc-finger transcription factor which binds to the two mouse type-I collagen gene promoters.
We have used the yeast two-hybrid system to clone the protein that interacts with the BFCOL1 (binding factor of a type-I collagen promoter) zinc-finger transcription factor that was cloned previously as the factor that binds to the two mouse proximal promoters of the type-I collagen genes. We utilized as bait the N-terminal domain of BFCOL1 that includes the zinc-finger DNA-binding domain. One cDNA contained a potential open reading frame for a polypeptide of 392 amino acids and was identical to PTRF (polymerase I and transcript-release factor), which is involved in transcription termination of the RNA polymerase I reaction. Northern-blot analysis revealed that the pattern of mRNA expression was similar to that of the type-I collagen gene. In addition, we detected the mRNA expression only in a fibroblast cell line and two bone cell lines, but not in other blood and neuronal cell lines. Recombinant protein was shown to enhance the binding of BFCOL1 to its binding site in the mouse proalpha2(I) collagen proximal promoter in vitro. The transient-transfection experiment showed that PTRF had a suppressive effect on the mouse proalpha2(I) collagen proximal promoter activity. We speculate that PTRF might play a role in the RNA polymerase II reaction as well as that of RNA polymerase I.
Hasegawa
T
T
Department of Basic Gerontology, National Institute for Longevity Sciences, 36-3, Gengo Morioka-cho, Obu, Aichi, 474-8522 Japan.
Takeuchi
A
A
Miyaishi
O
O
Xiao
H
H
Mao
J
J
Isobe
K
K
D000818:Animals; D001842:Bone and Bones / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics*; D004256:DNA Polymerase I / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins; D005347:Fibroblasts / Q000378:metabolism; D008565:Membrane Proteins; D051379:Mice; D016366:Open Reading Frames; D011347:Procollagen / Q000235:genetics; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D016335:Zinc Fingers
C112745:Cavin1 protein, mouse; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D011347:Procollagen; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors; C491741:Zfp148 protein, mouse; D003094:Collagen; D004256:DNA Polymerase I