Cloning and sequencing of human LHhCG receptor cDNA

Minegish, Takashi; Nakamura, Kazuto; Takakura, Yumi; Miyamoto, Kaoru; Hasegawa, Yoshihisa; Ibuki, Yoshito; Igarashi, Masao

doi:10.1016/0006-291x(90)91552-4

Cited by 237 publications

(86 citation statements)

References 11 publications

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“…1), spanning over a part of the signal peptide plus the entire extracellular, transmembrane and intracellular domains of the eLH/CG-R. The cDNA sequence (excluding primer sequence) of eLH/CG-R A (accession number AY464091 to GenBank) showed 92.3, 91.6, 88 and 85.7% homologies with reported cDNA sequences of porcine (Loosfelt et al 1989), bovine (Lussier et al 1996), human (Minegishi et al 1990) and rat (McFarland et al 1989) LH/CG-R respectively, and showed 64.6% homology with the equine follicle-stimulating hormone (FSH) receptor (FSH-R) cDNA sequence (Robert et al 1994). The deduced 680-amino acids sequence of eLH/CG-R A cDNA was similarly conserved among mammals (pig 92.5%, cow 91.6%, human 89.1% and rat 87.4%) and showed 58% homology with the equine FSH-R. Alignment with the porcine sequence suggests a putative cleavage site of the signal peptide between Gly 8 and Ala 9 (Loosfelt et al 1989).…”

Section: Nucleotide Sequence Analysis Of Elh/cg-r Cdna Isoformsmentioning

confidence: 78%

“…A 646-base pair (bp) cDNA fragment encoding a part of the extracellular domain of the LH/CG-receptor was first generated using primer pair P1 and P2 based on cDNA sequences similar among the bovine (Lussier et al 1996), porcine (Loosfelt et al 1989), human (Minegishi et al 1990) and murine (McFarland et al 1989) LH/CG-Rs (Table 1). First-strand cDNA was subjected to 30 cycles of PCR amplification using Advantage 2 polymerase mix (BD Biosciences, Palo Alto, CA, USA) and primer pair P1 and P2.…”

Section: Isolation and Characterization Of Elh/cg-r Cdna Isoformsmentioning

confidence: 99%

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Expression of the full-length and alternatively spliced equine luteinizing hormone/chorionic gonadotropin receptor mRNAs in the primary corpus luteum and fetal gonads during pregnancy

Saint‐Dizier

Chopineau²,

Dupont³

et al. 2004

Reproduction

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The full-length equine luteinizing hormone/chorionic gonadotropin (LH/CG) receptor (eLH/CG-R A ) cDNA and two alternatively spliced isoforms (eLH/CG-R B,C ) were isolated from luteal tissue and characterized using a combination of reverse transcription-polymerase chain reaction (RT-PCR) and 5 0 -rapid amplification of cDNA ends. The 680-amino acid full sequence of eLH/CG-R A displayed 87 -92% homology with other mammalian LH/CG-Rs. The eLH/CG-R B and eLH/CG-R C cDNA isoforms were truncated from the 3 0 -end of exon X: eLH/CG-R B spliced out of frame into the last exon whereas eLH/CG-R C contained an in-frame stop codon within a divergent sequence. Consequently, both eLH/CG-R B and eLH/CG-R C cDNA isoforms encoded putative proteins without transmembrane and intracellular domains. In order to study the responsiveness of the primary corpus luteum (CL) and fetal gonads to eCG, the expression of eLH/CG-R mRNAs was examined by RT-PCR and Northern blot analysis during early and mid-pregnancy. All three eLH/CG-R cDNA isoforms (eLH/CG-R A,B,C ) were expressed from day 14 to day 83 of pregnancy in the primary CL and from day 44 to day 222 in fetal gonads. Interestingly, the primary CL at days 89 and 151 expressed only truncated eLH/CG-R cDNA isoforms. The relative values of Northern hybridized major 7, 5.7, 3.9 and 1.8 kb eLH/CG-R mRNA transcripts tended to decrease in the primary CL whereas the unique major 1.8 kb eLH/CG-R mRNA was steadily expressed in fetal gonads during pregnancy. These results show that the expression of eLH/CG-R mRNAs occurs in the fetal gonads before ceasing in the primary CL and suggest that eCG may be involved in the gradual transition from a luteal to a feto-placental output of steroids during equine pregnancy.

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Section: Nucleotide Sequence Analysis Of Elh/cg-r Cdna Isoformsmentioning

confidence: 78%

Section: Isolation and Characterization Of Elh/cg-r Cdna Isoformsmentioning

confidence: 99%

Expression of the full-length and alternatively spliced equine luteinizing hormone/chorionic gonadotropin receptor mRNAs in the primary corpus luteum and fetal gonads during pregnancy

Saint‐Dizier

Chopineau²,

Dupont³

et al. 2004

Reproduction

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“…O LH e o hCG exercem seus efeitos biológicos através de um único receptor localizado na membrana citoplasmática (4). O receptor maduro do LH/hCG é um polipeptídeo de 674 aminoácidos, caracterizado por uma estrutura composta de três domínios: o extenso domínio extracelular aminoterminal, o domínio intermediário composto de sete segmentos helicoidais de propriedade hidrofóbica, interligados por 3 alças extracelulares e 3 alças intracelulares, e o curto domínio intracelular carboxi-terminal (4,5).…”

Section: O Receptor Do Lhunclassified

“…O receptor maduro do LH/hCG é um polipeptídeo de 674 aminoácidos, caracterizado por uma estrutura composta de três domínios: o extenso domínio extracelular aminoterminal, o domínio intermediário composto de sete segmentos helicoidais de propriedade hidrofóbica, interligados por 3 alças extracelulares e 3 alças intracelulares, e o curto domínio intracelular carboxi-terminal (4,5). Aproximadamente metade dos aminoácidos do receptor do LH está presente no domínio hidrofílico extracelular aminoterminal.…”

Section: O Receptor Do Lhunclassified

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Mutações ativadoras do gene do receptor do hormônio luteinizante em meninos com testotoxicose

Latrônico

2001

Arq Bras Endocrinol Metab

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RESUMOA testotoxicose é uma forma rara de puberdade precoce familial em meninos com herança autossômica dominante. Os caracteres sexuais secundários ocorrem geralmente antes dos 4 anos de idade. Nesta condição, níveis puberais de testosterona estão associados a níveis suprimidos ou pré puberais de gonadotrofinas. ABSTRACTTestotoxicosis is a rare form of familial precocious puberty in boys with autossomal dominant inheritance. Secondary sexual features usually occur before 4 years of age. In this condition, testosterone are elevated with suppressed or prepubertal gonadotropin levels. Several germline activating mutations in exon 11 of the LH receptor gene have been described in boys with testotoxicosis. The molecular analysis of 8 Brazilian boys with testotoxicosis revealed 5 different mutations, 3 of them identified exclusively in Brazil: Ala568Val, Leu457Arg, and Leu368Pro, located in the third intracellular loop and in the III and I transmembrane helices, respectively. The Ala568Val mutation was found in 42.8% of the Brazilian families. Women with activating mutations, mother or sisters of boys with testotoxicosis, did not develop precocious puberty and showed normal reproductive function. Two Brazilian women, including a prepubertal girl with activating mutations, were asymptomatic and had normal hormonal profile. Somatic activating mutations of the LH receptor gene were recently identified in 3 boys with Leydig cell tumors. However, a recent report did not find such mutations in 4 Leydig tumors, 3 tecomas and 4

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Detection of metastatic breast cancer by β-hCG polymerase chain reaction

et al. 1996

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Reverse transcriptase-polymerase chain reaction (RT-PCR) for detection of occult malignancies in breast cancer patients is evolving as a useful diagnostic tool. However, no reliable molecular mRNA markers are available. We developed an RT-PCR plus Southern blot assay using p-hCG @-subunit of human chorionic gonadotropin) gene expression as a tumor marker for detection of breast malignancies metastatic to tumor-draining lymph nodes and blood. Breast carcinoma cell lines, primary breast malignancies and human placenta were used as positive controls for establishing the P-hCG RT-PCR assay. Peripheral blood leukocytes (PBL) from normal volunteer donors, normal breast tissue and lymph nodes from cancer-free patients were used as negative controls. P-hCG RT-PCR was used to assess tumor cell presence in PBL and tumor-draining axillary nodes from patients with AJCC stage I-IV breast cancer. The assay sensitivity and specificity were enhanced by restriction endonuclease digestion of an Sty I site of the RT-PCR cDNA product followed by Southern blot analysis. P-hCG mRNA was expressed in all breast cancer cell lines and 80% of primary breast cancers; it was not expressed in negative Neoplastic cells aberrantly express genes that are silent in their normal counterparts. These genes often encode for proteins that endow neoplastic cells selective advantages with respect to growth and other malignant cell properties, such as metastasis and invasion. Neoplastic cells produce autocrine growth factors or hormones that are not produced or are expressed at low levels by normal cells. Human chorionic gonadotropin (hCG) is a hormone produced by the trophoblast and is critical to sustaining pregnancy (Moyle and Campbell, 1993). hCG and its individual subunits are produced not only in gestational trophoblastic and germ-cell cancers but also in a wide range of non-gonadal tumors, such as gastrointestinal, lung and breast carcinomas (McManus et al., 1976; Madersbacher et al., 1994; Marcillac et al., 1992;Acevedo et al., 1995). We have demonstrated its usefulness as a molecular marker for melanoma (Doi et al., 1996). The pathophysiologic role for the expression of hCG in malignant neoplasms is currently speculative . hCG expression represents recapitulation of fetal cell properties by malignant cells.Immunohistochemical analysis using polyclonal or monoclonal antibodies (MAbs) has demonstrated hCG as a cancer marker (Krichevsky et al., 1995). However, these studies were not always specific, primarily due to the recognition of crossreactive epitopes or subunit chains of related gonadotropins (McManus et al., 1976; Krichevsky et al., 1995). About 7040% of adenocarcinomas arising in the mammary gland stain for the p subunit of hCG (P-hCG) by immunohistochemical techniques (Agnantis et al., 1992). hCG is a dimeric hormone composed of 2 non-covalently linked subunits designated a and P (Bo and Boime, 1992;Boorstein et al., 1982). It belongs to a glycoprotein hormone family, sharing many similarities with luteinizing hormone (LH), follicle stim...

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Cloning and sequencing of human LHhCG receptor cDNA

Cited by 237 publications

References 11 publications

Expression of the full-length and alternatively spliced equine luteinizing hormone/chorionic gonadotropin receptor mRNAs in the primary corpus luteum and fetal gonads during pregnancy

Expression of the full-length and alternatively spliced equine luteinizing hormone/chorionic gonadotropin receptor mRNAs in the primary corpus luteum and fetal gonads during pregnancy

Mutações ativadoras do gene do receptor do hormônio luteinizante em meninos com testotoxicose

Detection of metastatic breast cancer by β-hCG polymerase chain reaction

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