Ala/Thr201 in extracellular loop 2 and Leu/Phe290 in transmembrane domain 6 of type 1 frog gonadotropin-releasing hormone receptor confer differential ligand sensitivity and signal transduction

Jae Young Seong, Li Wang, Da Young Oh, Oim Yun, Kaushik Maiti, Jian Hua Li, Jae Mok Soh, Hueng Sik Choi, Kyungjin Kim, Hubert Vaudry, Hyuk Bang Kwon

Research output: Contribution to journalArticle

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Abstract

Recently, we have identified three distinct types of bullfrog GnRH receptor (designated bfGnRHR-1, bfGnRHR-2, and bfGnRHR-3). In the present study, we have isolated three GnRHR clones in Rana dybowskii (dyGnRHR-1, dyGnRHR-2, and dyGnRHR-3). Despite high homology of dyGnRHRs with the corresponding bfGnRHRs, dyGnRHRs revealed different signaling pathways and ligand sensitivity compared with the bfGnRHR counterparts. Activation of dyGnRHRs with GnRH stimulated cAMP-mediated gene expression. However, dyGnRHR-3 but not dyGnRHR-1 and -2 induced c-fos promoter-driven gene expression. Consistently, dyGnRHR-1 and dyGnRHR-2 were not able to increase GnRH-induced inositol phosphate accumulation, whereas all bfGnRHRs and dyGnRHR-3 were, indicating that dyGnRHR-1 and dyGnRHR-2 are coupled to solely Gs, whereas all bfGnRHRs and dyGnRHR-3 are coupled to both Gs and Gq/11. Moreover, dyGnRHR-1 and dyGnRHR-2 showed about 10-fold less sensitivity to each ligand than that of the bfGnRHR counterparts. Using type 1 chimeric and point-mutated receptors, we further elucidated that specific amino acids, Ala/Thr201 in extracellular loop 2 and Leu/Phe290 in transmembrane domain 6 of the type 1 receptor, are responsible for ligand sensitivity and signal transduction pathway. Particularly, substitution of Leu290 to Phe in dyGnRHR-1 increased GnRH-induced inositol phosphate production as well as c-fos promoter-driven gene expression whereas substitution of Phe290 to Leu in bfGnRHR-1 decreased those activities. Collectively, these results demonstrate the presence of three types of GnRHR in amphibians, and suggest species- and type-specific ligand recognition and different signaling pathways in frog GnRHRs.

Original languageEnglish
Pages (from-to)454-466
Number of pages13
JournalEndocrinology
Volume144
Issue number2
DOIs
Publication statusPublished - 2003 Feb 1
Externally publishedYes

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LHRH Receptors
Anura
Signal Transduction
Gonadotropin-Releasing Hormone
Ligands
Inositol Phosphates
Gene Expression
Ranidae
Rana catesbeiana
Amphibians
Clone Cells
Amino Acids

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Ala/Thr201 in extracellular loop 2 and Leu/Phe290 in transmembrane domain 6 of type 1 frog gonadotropin-releasing hormone receptor confer differential ligand sensitivity and signal transduction. / Seong, Jae Young; Wang, Li; Oh, Da Young; Yun, Oim; Maiti, Kaushik; Li, Jian Hua; Soh, Jae Mok; Choi, Hueng Sik; Kim, Kyungjin; Vaudry, Hubert; Kwon, Hyuk Bang.

In: Endocrinology, Vol. 144, No. 2, 01.02.2003, p. 454-466.

Research output: Contribution to journalArticle

Seong, Jae Young ; Wang, Li ; Oh, Da Young ; Yun, Oim ; Maiti, Kaushik ; Li, Jian Hua ; Soh, Jae Mok ; Choi, Hueng Sik ; Kim, Kyungjin ; Vaudry, Hubert ; Kwon, Hyuk Bang. / Ala/Thr201 in extracellular loop 2 and Leu/Phe290 in transmembrane domain 6 of type 1 frog gonadotropin-releasing hormone receptor confer differential ligand sensitivity and signal transduction. In: Endocrinology. 2003 ; Vol. 144, No. 2. pp. 454-466.
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abstract = "Recently, we have identified three distinct types of bullfrog GnRH receptor (designated bfGnRHR-1, bfGnRHR-2, and bfGnRHR-3). In the present study, we have isolated three GnRHR clones in Rana dybowskii (dyGnRHR-1, dyGnRHR-2, and dyGnRHR-3). Despite high homology of dyGnRHRs with the corresponding bfGnRHRs, dyGnRHRs revealed different signaling pathways and ligand sensitivity compared with the bfGnRHR counterparts. Activation of dyGnRHRs with GnRH stimulated cAMP-mediated gene expression. However, dyGnRHR-3 but not dyGnRHR-1 and -2 induced c-fos promoter-driven gene expression. Consistently, dyGnRHR-1 and dyGnRHR-2 were not able to increase GnRH-induced inositol phosphate accumulation, whereas all bfGnRHRs and dyGnRHR-3 were, indicating that dyGnRHR-1 and dyGnRHR-2 are coupled to solely Gs, whereas all bfGnRHRs and dyGnRHR-3 are coupled to both Gs and Gq/11. Moreover, dyGnRHR-1 and dyGnRHR-2 showed about 10-fold less sensitivity to each ligand than that of the bfGnRHR counterparts. Using type 1 chimeric and point-mutated receptors, we further elucidated that specific amino acids, Ala/Thr201 in extracellular loop 2 and Leu/Phe290 in transmembrane domain 6 of the type 1 receptor, are responsible for ligand sensitivity and signal transduction pathway. Particularly, substitution of Leu290 to Phe in dyGnRHR-1 increased GnRH-induced inositol phosphate production as well as c-fos promoter-driven gene expression whereas substitution of Phe290 to Leu in bfGnRHR-1 decreased those activities. Collectively, these results demonstrate the presence of three types of GnRHR in amphibians, and suggest species- and type-specific ligand recognition and different signaling pathways in frog GnRHRs.",
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T1 - Ala/Thr201 in extracellular loop 2 and Leu/Phe290 in transmembrane domain 6 of type 1 frog gonadotropin-releasing hormone receptor confer differential ligand sensitivity and signal transduction

AU - Seong, Jae Young

AU - Wang, Li

AU - Oh, Da Young

AU - Yun, Oim

AU - Maiti, Kaushik

AU - Li, Jian Hua

AU - Soh, Jae Mok

AU - Choi, Hueng Sik

AU - Kim, Kyungjin

AU - Vaudry, Hubert

AU - Kwon, Hyuk Bang

PY - 2003/2/1

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N2 - Recently, we have identified three distinct types of bullfrog GnRH receptor (designated bfGnRHR-1, bfGnRHR-2, and bfGnRHR-3). In the present study, we have isolated three GnRHR clones in Rana dybowskii (dyGnRHR-1, dyGnRHR-2, and dyGnRHR-3). Despite high homology of dyGnRHRs with the corresponding bfGnRHRs, dyGnRHRs revealed different signaling pathways and ligand sensitivity compared with the bfGnRHR counterparts. Activation of dyGnRHRs with GnRH stimulated cAMP-mediated gene expression. However, dyGnRHR-3 but not dyGnRHR-1 and -2 induced c-fos promoter-driven gene expression. Consistently, dyGnRHR-1 and dyGnRHR-2 were not able to increase GnRH-induced inositol phosphate accumulation, whereas all bfGnRHRs and dyGnRHR-3 were, indicating that dyGnRHR-1 and dyGnRHR-2 are coupled to solely Gs, whereas all bfGnRHRs and dyGnRHR-3 are coupled to both Gs and Gq/11. Moreover, dyGnRHR-1 and dyGnRHR-2 showed about 10-fold less sensitivity to each ligand than that of the bfGnRHR counterparts. Using type 1 chimeric and point-mutated receptors, we further elucidated that specific amino acids, Ala/Thr201 in extracellular loop 2 and Leu/Phe290 in transmembrane domain 6 of the type 1 receptor, are responsible for ligand sensitivity and signal transduction pathway. Particularly, substitution of Leu290 to Phe in dyGnRHR-1 increased GnRH-induced inositol phosphate production as well as c-fos promoter-driven gene expression whereas substitution of Phe290 to Leu in bfGnRHR-1 decreased those activities. Collectively, these results demonstrate the presence of three types of GnRHR in amphibians, and suggest species- and type-specific ligand recognition and different signaling pathways in frog GnRHRs.

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