The chromosomal gene structure and two mRNAs for human granulocyte colony-stimulating factor.
Two different cDNAs for human granulocyte colony-stimulating factor (G-CSF) were isolated from a cDNA library constructed with mRNA prepared from human squamous carcinoma cells, which produce G-CSF constitutively. The nucleotide sequence analysis of both cDNAs indicated that two polypeptides coded by these cDNAs are different at one position where three amino acids are deleted/inserted. When the two cDNAs were introduced into monkey COS cells under the SV40 early promoter, both of them produced proteins having authentic G-CSF activity and some difference in the specific activity was suggested. A human gene library was then screened with the G-CSF cDNA and the DNA fragment containing the G-CSF chromosomal gene was characterized by the nucleotide sequence analysis. The human G-CSF gene is interrupted by four introns and a comparison of the structures of the two G-CSF cDNAs with that of the chromosomal gene indicated that the two mRNAs are generated by alternative use of two 5' splice donor sequences in the second intron of the G-CSF gene. When the G-CSF chromosomal gene was expressed in monkey COS cells by using the SV40 enhancer two mRNAs were detected by S1 mapping analysis.
Nagata
S
S
Tsuchiya
M
M
Asano
S
S
Yamamoto
O
O
Hirata
Y
Y
Kubota
N
N
Oheda
M
M
Nomura
H
H
Yamazaki
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002294:Carcinoma, Squamous Cell; D002460:Cell Line; D002522:Chlorocebus aethiops; D002874:Chromosome Mapping; D002877:Chromosomes, Human / Q000502:physiology; D003001:Cloning, Molecular; D003115:Colony-Stimulating Factors / Q000235:genetics*; D004247:DNA / Q000302:isolation & purification; D004262:DNA Restriction Enzymes; D005796:Genes; D016179:Granulocyte Colony-Stimulating Factor; D006098:Granulocytes / Q000166:cytology*; D006801:Humans; D007668:Kidney; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics*
D003115:Colony-Stimulating Factors; D012333:RNA, Messenger; D016179:Granulocyte Colony-Stimulating Factor; D004247:DNA; D004262:DNA Restriction Enzymes