Alternate splicing of mRNAs encoding human mast cell growth factor and localization of the gene to chromosome 12q22-q24.
Human mast cell growth factor (MGF) complementary DNAs (cDNAs) were cloned from HeLa cells using the polymerase chain reaction with oligonucleotides corresponding to murine and human MGF sequences. Sequencing of the cloned human MGF polymerase chain reaction products revealed two types of cDNA: a full length form corresponding in size to the murine cDNA, and an alternately spliced clone with a deletion of the sixth exon of the gene. Since membrane-bound MGF is predicted to be proteolytically cleaved within the sequences encoded by exon 6 to generate a soluble protein, this alternately spliced cDNA would likely encode a noncleavable, membrane-bound form of MGF. No difference in biological activity on human bone marrow cells was observed with recombinant, soluble forms of both types of human MGF protein. Our previous localization of the murine MGF gene to the Sl locus on chromosome 10 suggested (via conserved linkage groups) that the human MGF gene would be located on human chromosome 12. Therefore, rodent-human somatic cell hybrids with or without an entire human chromosome 12 and hybrids retaining partial 12 were tested by Southern blot analysis and used to show the presence of the human Mgf locus at chromosome region 12q. Chromosomal in situ hybridization localized the gene to 12q22-q24 in the region predicted by the comparative mapping of the murine Mgf/Sl locus.
Anderson
D M
DM
Department of Molecular Biology, Immunex Research and Development Corporation, Seattle, Washington 98101.
Williams
D E
DE
Tushinski
R
R
Gimpel
S
S
Eisenman
J
J
Cannizzaro
L A
LA
Aronson
M
M
Croce
C M
CM
Huebner
K
K
Cosman
D
D
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D002874:Chromosome Mapping; D002881:Chromosomes, Human, Pair 12; D003001:Cloning, Molecular; D004357:Drug Synergism; D004921:Erythropoietin / Q000494:pharmacology; D005091:Exons / Q000235:genetics; D015870:Gene Expression / Q000502:physiology; D006367:HeLa Cells; D016298:Hematopoietic Cell Growth Factors / Q000235:genetics* / Q000494:pharmacology; D006801:Humans; D006822:Hybrid Cells; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012326:RNA Splicing / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology; D019089:Stem Cell Factor
D016298:Hematopoietic Cell Growth Factors; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D004921:Erythropoietin