Recharacterization of the start sites for the major human transforming growth factor-beta 1 mRNA.
The start sites for the major human TGF-beta 1 transcripts have been reexamined. A comparison of ribonuclease and S1 nuclease protection analyses on native TGF-beta 1 mRNA and in vitro transcribed human TGF-beta 1 transcripts of defined sizes places the most 5' start site for the native TGF-beta 1 message approx. 50 nucleotides upstream from the previously published start site at base +1. Furthermore, the same techniques indicate that the apparent downstream start site at base +271 is an artefact due to the presence of an A + T-rich island in the middle of an otherwise highly G + C-rich sequence. This is not apparent if S1 nuclease protection is used alone, which emphasizes the importance of using the two techniques in combination for this type of analysis. Thus the major 2.5 kb TGF-beta 1 band seen on Northern blots comprises only mRNA transcribed from the more upstream of the two previously characterized promoters. This has important implications both for the transcriptional and translational regulation of this growth factor.
Romeo
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D
Laboratory of Chemoprevention, National Cancer Institute, Bethesda, MD 20892-5055, USA.
Allison
R S
RS
Kondaiah
P
P
Wakefield
L M
LM
D001482:Base Composition; D001943:Breast Neoplasms / Q000737:chemistry / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction; D015347:RNA Probes; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D012260:Ribonucleases / Q000032:analysis; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000032:analysis; D014158:Transcription, Genetic; D016212:Transforming Growth Factor beta / Q000235:genetics*; D014407:Tumor Cells, Cultured
D015347:RNA Probes; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D012260:Ribonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases