title: Emergence of an Ancestral Glycoprotein Hormone in the Pituitary of the Sea Lamprey, a Basal Vertebrate.
abstract: The gnathostome (jawed vertebrates) classical pituitary glycoprotein hormones, FSH, LH, and TSH, consist of a common α-subunit (GpA1) and unique β-subunits (Gpβ1, -2, and -3), whereas a recently identified pituitary glycoprotein hormone, thyrostimulin, consists of GpA2 and GpB5. This paper reports the identification, expression, and function of an ancestral, nonclassical, pituitary heterodimeric glycoprotein hormone (GpH) consisting of the thyrostimulin A2 subunit with the classical β-subunit in the sea lamprey, Petromyzon marinus, a jawless basal vertebrate. Lamprey (l) GpA2, and lGpHβ were shown to form a heterodimer by coimmunoprecipitation of lGpA2 with FLAG-tagged lGpHβ after the overexpression in transiently transfected COS7 cells using a bipromoter vector. Dual-label fluorescent in situ hybridization and immunohistochemistry showed the coexpression of individual subunits in the proximal pars distalis of the pituitary. GnRH-III (1μΜ) significantly increased the expression of lGpHβ and lGpA2 in in vitro pituitary culture. Recombinant lamprey GpH was constructed by tethering the N terminal of lGpA2 to the C terminal of lGpHβ with a linker region composed of six histidine residues followed by three glycine-serine repeats. This recombinant lamprey GpH activated the lamprey glycoprotein hormone receptor I as measured by increased cAMP/luciferase activity. These data are the first to demonstrate a functional, unique glycoprotein heterodimer that is not found in any other vertebrate. These data suggest an intermediate stage of the structure-function of the gonadotropin/thyroid-stimulating hormone in a basal vertebrate, leading to the emergence of the highly specialized gonadotropin hormones and thyroid stimulating hormones in gnathostomes.
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Stacia A
authors.initials: SA
authors.lastname: Sower
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Wayne A
authors.initials: WA
authors.lastname: Decatur
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Krist N
authors.initials: KN
authors.lastname: Hausken
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Marquis
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Shannon L
authors.initials: SL
authors.lastname: Barton
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: James
authors.initials: J
authors.lastname: Gargan
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Mihael
authors.initials: M
authors.lastname: Freamat
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Michael
authors.initials: M
authors.lastname: Wilmot
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Lian
authors.initials: L
authors.lastname: Hollander
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Hall
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Masumi
authors.initials: M
authors.lastname: Nozaki
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Michal
authors.initials: M
authors.lastname: Shpilman
authors.affiliation: Center for Molecular and Comparative Endocrinology (C.A.S., W.A.D., K.N.H., T.J.M., S.L.B., J.G., M.F., M.W., J.A.H.), Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824; Robert H. Smith Faculty of Agriculture, Food, and Environment (L.H., M.S., B.L.-S.), The Hebrew University of Jerusalem, Rehovot, Israel 76100; and Sado Marine Biological Station (M.N.), Niigata University, Tassha, Sado, Niigata 952-2135, Japan.
authors.forename: Berta
authors.initials: B
authors.lastname: Levavi-Sivan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D019143:Evolution, Molecular; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D007798:Lampreys / Q000235:genetics*; D008969:Molecular Sequence Data; D036361:Peptide Hormones / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D010902:Pituitary Gland / Q000378:metabolism; D010907:Pituitary Hormones / Q000235:genetics* / Q000378:metabolism; D017385:Sequence Homology; D014714:Vertebrates / Q000145:classification / Q000235:genetics*
chemical_list: D006023:Glycoproteins; D036361:Peptide Hormones; D010907:Pituitary Hormones