An alternatively processed mRNA generated from human hepatocyte growth factor gene.
We recently reported the isolation and sequencing of cDNA for human hepatocyte growth factor (hHGF) [Miyazawa, K., Tsubouchi, H., Naka, D., Takahashi, K., Okigaki, M., Arakaki, N., Nakayama, H., Hirono, S., Sakiyama, O., Takahashi, K., Gohda, E., Daikuhara, Y. & Kitamura, N. (1989) Biochem. Biophys. Res. Commun. 163, 967-973]. In the present study, we report the sequence of another cDNA clone for a shorter form of hHGF mRNA. Comparison of the sequence with that of the hHGF cDNA revealed that the two sequences are identical in their 5' ends up to 865 nucleotides downstream from the translation-initiation site, then completely diverge from each other. By Northern blot analysis, the hHGF-related 1.5-kb mRNA, which corresponded to the newly isolated cDNA variant, was identified in human placenta. Sequence analysis of a human genomic HGF clone showed that the diverged 3'-terminal portion of the mRNA is generated by alternative RNA-processing events utilizing a specific exon. The mRNA could encode a short hHGF molecule of 290 amino acids corresponding to the N-terminal portion of hHGF which consists of 728 amino acids. In order to examine the effect of the predicted translation product on hepatocyte growth, an expression plasmid for the cDNA variant was constructed and transfected into Cos cells. Immunoblotting analysis showed that the transfected Cos cells produced a protein of about 33 kDa. The protein product did not stimulate DNA synthesis by rat hepatocytes in primary culture.
Miyazawa
K
K
Institute for Liver Research, Kansai Medical University, Moriguchi, Japan.
Kitamura
A
A
Naka
D
D
Kitamura
N
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004261:DNA Replication; D005091:Exons; D005796:Genes; D006133:Growth Substances / Q000235:genetics*; D017228:Hepatocyte Growth Factor; D006801:Humans; D007438:Introns; D008099:Liver / Q000502:physiology*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics* / Q000302:isolation & purification; D051381:Rats; D015183:Restriction Mapping; D014162:Transfection
D006133:Growth Substances; D012333:RNA, Messenger; D017228:Hepatocyte Growth Factor; D004247:DNA