Characterization of Three Abundant mRNAs from Human Ovarian Granulosa Cells

Abstract

Three cDNA clones, pHGR122, pHGR11, and pHGR74 containing the coding information for abundant mRNAs were identified from a human ovarian granulosa cell cDNA library. Characterization by nucleotide sequencing revealed that pHGR122 was specific for a collagenase inhibitor and pHGR11 for melanoma-associated antigen ME491. Relative quantification by Northern analysis indicated that collagenase inhibitor mRNA is a major species in granulosa cells. This finding provides evidence for the origin of this protein in follicular fluid as a secretory product of granulosa cells. pHGR11 identified melanoma-associated antigen ME491 as the unexpected product of normal, noncarcinogenic, granulosa cells. pHGR74 has the complete coding information for an unknown protein. Three independent experiments: (i) cell-free translation of pHGR74 RNA; (ii) transcription of suitable restriction fragments followed by cell-free translation; (iii) hydrolysis of the cell-free translation product of pHGR74 RNA by endoproteinase Lys-C, identified one open reading frame coding for an acidic, highly hydrophilic protein of 111 amino acid residues. pHGR74 mRNA is expressed in human testis, prostate, seminal vesicle, and ovarian granulosa cells. A comparative Southern analysis indicates pHGR74 mRNA is species specific and encoded by a single-copy gene.