Identification and characterization of a novel member of olfactomedin-related protein family, hGC-1, expressed during myeloid lineage development.
We have cloned a novel hematopoietic granulocyte colony-stimulating factor (G-CSF)-induced olfactomedin-related glycoprotein, termed hGC-1 (human G-CSF-stimulated clone-1). mRNA differential display was used in conjunction with a modified two-phase liquid culture system. Cultures were enriched for early precursors of erythroid, myeloid, and megakaryocytic lineages, which were isolated after induction with erythropoietin, G-CSF, and thrombopoietin, respectively. RNA from the enriched cells was subjected to differential display analysis to identify lineage-specific expressed genes. One clone specifically induced by G-CSF, hGC-1, was characterized. The 2861 bp cDNA clone of hGC-1 contained an open reading frame of 1530 nucleotides, translating into a protein of 510 amino acids with a signal peptide and six N-linked glycosylation motifs. The protein sequence of hGC-1 showed it to be a glycoprotein of the olfactomedin family, which includes olfactomedin, TIGR, Noelin-2 and latrophilin-1. Olfactomedin-like genes show characteristic tissue-restricted patterns of expression; the specific tissues expressing these genes differ among the family members. hGC-1 was strongly expressed in the prostate, small intestine, and colon, moderately expressed in the bone marrow and stomach, and not detectable in other tissues. In vitro translation and ex vivo expression showed hGC-1 to be an N-linked glycoprotein. The hGC-1 gene locus mapped to chromosome 13q14.3. Together, our findings indicate that hGC-1 is primarily expressed as an extracellular olfactomedin-related glycoprotein during normal myeloid-specific lineage differentiation, suggesting the possibility of a matrix-related function for hGC-1 in differentiation.
Zhang
Jiachang
J
Molecular and Clinical Hematology Branch, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Liu
Wen Li
WL
Tang
Delia C
DC
Chen
Ling
L
Wang
Min
M
Pack
Svetlana D
SD
Zhuang
Zhengping
Z
Rodgers
Griffin P
GP
D000581:Amidohydrolases / Q000378:metabolism; D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000494:pharmacology; D001798:Blood Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D018826:CD13 Antigens / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002874:Chromosome Mapping; D002882:Chromosomes, Human, Pair 13 / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006021:Glycophorins / Q000494:pharmacology; D006023:Glycoproteins / Q000235:genetics*; D006031:Glycosylation; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics* / Q000378:metabolism; D018922:HL-60 Cells; D006410:Hematopoiesis / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D039661:Integrin beta3; D020014:K562 Cells; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D022423:Myeloid Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D010980:Platelet Membrane Glycoproteins / Q000494:pharmacology; D014176:Protein Biosynthesis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D001798:Blood Proteins; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D006021:Glycophorins; D006023:Glycoproteins; D039661:Integrin beta3; C454127:OLFM4 protein, human; D010980:Platelet Membrane Glycoproteins; C081451:olfactomedin; D016179:Granulocyte Colony-Stimulating Factor; D018826:CD13 Antigens; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase