Identification of Alternatively Spliced Transcripts Encoding Murine Macrophage Colony-Stimulating Factor
Identifieur interne : 003B61 ( Main/Exploration ); précédent : 003B60; suivant : 003B62Identification of Alternatively Spliced Transcripts Encoding Murine Macrophage Colony-Stimulating Factor
Auteurs : Shinya Suzu [Japon] ; Kiyohiko Hatake [Japon] ; Jun Ota [Japon] ; Yuji Mishima [Japon] ; Muneo Yamada [Japon] ; Seiichi Shimamura [Japon] ; Fumihiko Kimura [Japon] ; Kazuo Motoyoshi [Japon]Source :
- Biochemical and Biophysical Research Communications [ 0006-291X ] ; 1998.
English descriptors
- Teeft :
- Active protein, Alternate initiation codon, Amino, Amino acid residues, Amino acid sequence, Bbrc, Bbrcg, Biol, Biophysical research communications, Broblastic cell line, Cdna, Cdna fragment, Cdna inserts, Codon, Complete exon, Cos7, Cos7 cells, Entire coding region, Ethidium bromide staining, Exon, Human gene, Initiation codon, Insertion, Insertion sequence, Intron, Intron sequence, Mature protein, Mcsf transcript, Motoyoshi, Murine, Northern blot analysis, Nucleic acids, Nucleotide, Nucleotide sequences, Plasmid, Possible acceptor sequence, Primer, Primer pair, Reading frame, Sequencing, Signal sequence, Stromal, Suzu, Takara shuzo, Transcript.
Abstract
Abstract: We have isolated a novel cDNA encoding macrophage colony-stimulating factor (M-CSF) from a murine stromal cell line, ST2. The cDNA included an entire coding sequence of the M-CSF gene but contained an additional sequence of 140 base pairs (bp). Northern blot analysis demonstrated that other murine cell lines such as a fibroblastic cell line (L) and a stromal cell line (PA6) also expressed the transcripts corresponding to the clone. The nucleotide sequence analyses of the cDNA and the cloned M-CSF genome revealed that the 140-bp insertion sequence was part of intron 1 which separated exon 1 and exon 2: the former contained part of the amino acid residues of the signal sequence and the latter the rest of the signal sequence and the first 22 amino acid residues of the mature protein. The insertion of the 140-bp intron sequence not only changed the amino acid sequence of the signal peptide but also generated an in-frame termination codon. However, instead of the dysfunction of the original initiation codon, the 140-bp insertion sequence contained a putative ATG initiation codon that preserved the original open reading frame. Finally, we found that the cDNA directed the expression of a secreted and biologically active M-CSF protein when it was introduced into COS7 cells and M-CSF activity in the culture supernatants was measured using an M-CSF-dependent cell line. These results indicate the presence of an alternatively spliced M-CSF transcript which utilizes an alternate initiation codon in order to specify active M-CSF protein.
Url:
DOI: 10.1006/bbrc.1998.8394
Affiliations:
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Le document en format XML
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The cDNA included an entire coding sequence of the M-CSF gene but contained an additional sequence of 140 base pairs (bp). Northern blot analysis demonstrated that other murine cell lines such as a fibroblastic cell line (L) and a stromal cell line (PA6) also expressed the transcripts corresponding to the clone. The nucleotide sequence analyses of the cDNA and the cloned M-CSF genome revealed that the 140-bp insertion sequence was part of intron 1 which separated exon 1 and exon 2: the former contained part of the amino acid residues of the signal sequence and the latter the rest of the signal sequence and the first 22 amino acid residues of the mature protein. The insertion of the 140-bp intron sequence not only changed the amino acid sequence of the signal peptide but also generated an in-frame termination codon. However, instead of the dysfunction of the original initiation codon, the 140-bp insertion sequence contained a putative ATG initiation codon that preserved the original open reading frame. Finally, we found that the cDNA directed the expression of a secreted and biologically active M-CSF protein when it was introduced into COS7 cells and M-CSF activity in the culture supernatants was measured using an M-CSF-dependent cell line. These results indicate the presence of an alternatively spliced M-CSF transcript which utilizes an alternate initiation codon in order to specify active M-CSF protein.</div></front></TEI><affiliations><list><country><li>Japon</li></country></list><tree><country name="Japon"><noRegion><name sortKey="Suzu, Shinya" sort="Suzu, Shinya" uniqKey="Suzu S" first="Shinya" last="Suzu">Shinya Suzu</name></noRegion><name sortKey="Hatake, Kiyohiko" sort="Hatake, Kiyohiko" uniqKey="Hatake K" first="Kiyohiko" last="Hatake">Kiyohiko Hatake</name><name sortKey="Kimura, Fumihiko" sort="Kimura, Fumihiko" uniqKey="Kimura F" first="Fumihiko" last="Kimura">Fumihiko Kimura</name><name sortKey="Mishima, Yuji" sort="Mishima, Yuji" uniqKey="Mishima Y" first="Yuji" last="Mishima">Yuji Mishima</name><name sortKey="Motoyoshi, Kazuo" sort="Motoyoshi, Kazuo" uniqKey="Motoyoshi K" first="Kazuo" last="Motoyoshi">Kazuo Motoyoshi</name><name sortKey="Ota, Jun" sort="Ota, Jun" uniqKey="Ota J" first="Jun" last="Ota">Jun Ota</name><name sortKey="Shimamura, Seiichi" sort="Shimamura, Seiichi" uniqKey="Shimamura S" first="Seiichi" last="Shimamura">Seiichi Shimamura</name><name sortKey="Yamada, Muneo" sort="Yamada, Muneo" uniqKey="Yamada M" first="Muneo" last="Yamada">Muneo Yamada</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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