Purification, cDNA cloning, and secretory properties of FLRG protein from PC12 cells and the distribution of FLRG mRNA and protein in rat tissues.
A 35 kD protein was isolated and purified from conditioned media of Bcl-2 cDNA-transfected PC12 cells and its cDNA cloned. A database analysis showed that the 35 kD protein is a rat homologue of the human FLRG protein. The biochemical as well as morphological properties of the rat FLRG protein in PC12 cells were examined and its distribution in rat tissues determined. The levels of FLRG mRNA expressed were low during the fetal period, compared with those of follistatin mRNA. The distribution of FLRG and follistatin mRNAs differed from each other after birth; the expression levels of FLRG mRNA were abundant in the adrenal gland and testis, whereas those of follistatin mRNA and activin A were markedly high in the ovary. The presence of FLRG mRNA and/or protein was confirmed in spermatocytes at various differentiating stages andin endocrine cells of both the adrenal cortex and medulla. When overexpressed in PC12 cells, the FLRG protein was found to be stored in secretory granules of the cells and largely secreted by a regulated pathway, while activin A enhancedthe constitutive secretion of the FLRG protein from wild-typpe PC12 cells, indicating that the FLRG protein possesses dualproperties in secretory pathways. The different distribution between FLRG and follistatin mRNA suggests that, like follistatin in the ovary, the FLRG protein may be involved in the maintenance of spermatogenesis in the testis and the growth and function of adrenal tissue cells, probably by regulating the functions of its binding partners such as the TGF-beta ( superfamily members.
Ohsawa
Yoshiyuki
Y
Department of Cell Biology and Neurosciences, Osaka University Graduated School of Medicine, Suita, Osaka, Japan.
Zhang
Guiqin
G
Kametaka
Satoshi
S
Shibata
Masahiro
M
Koike
Masato
M
Waguri
Satoshi
S
Uchiyama
Yasuo
Y
D028341:Activins; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D019992:Databases as Topic; D005260:Female; D038702:Follistatin-Related Proteins / Q000096:biosynthesis* / Q000235:genetics*; D015723:Gene Library; D015151:Immunoblotting; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D028322:Inhibin-beta Subunits; D008297:Male; D008969:Molecular Sequence Data; D016716:PC12 Cells; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism
D018076:DNA, Complementary; D038702:Follistatin-Related Proteins; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; C422405:activin A; D028341:Activins; D028322:Inhibin-beta Subunits