Regulation of the Guanylyl Cyclase-B Receptor by Alternative Splicing

Tamura, Naohisa; Garbers, David L.

doi:10.1074/jbc.m308680200

Cited by 37 publications

(32 citation statements)

References 48 publications

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“…1). Using specific primers that differentiated between previously characterised Npr2 splice variants (Tamura & Garbers 2003) we were able to detect expression of GC-B1, 2 and 3 in aT3-1 and primary pituitary, but abundance of the GC-B1 transcript in LbT2 cells was low, with no apparent GC-B2/3 expression. In addition, all cDNAs expressed soluble guanylyl cyclase Figure 1 Expression of natriuretic peptide components in immortalised gonadotroph cell lines and primary mouse pituitaries.…”

Section: Molecular Characterisation Of Gonadotroph Natriuretic Peptidmentioning

confidence: 85%

Molecular characterisation and functional interrogation of a local natriuretic peptide system in rodent pituitaries, αT3-1 and LβT2 gonadotroph cells

Thompson

Chand²,

Jonas

et al. 2009

Journal of Endocrinology

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In the pituitary, C-type natriuretic peptide (CNP) has been implicated as a gonadotroph-specific factor, yet expression of the CNP gene (Nppc) and CNP activity in gonadotrophs is poorly defined. Here, we examine the molecular expression and putative function of a local gonadotroph natriuretic peptide system. Nppc, along with all three natriuretic peptide receptors (Npr1, Npr2 and Npr3), was expressed in both aT3-1 and LbT2 cells and primary mouse pituitary tissue, yet the genes for atrial-(ANP) and B-type natriuretic peptides (Nppa and Nppb) were much less abundant. Putative processing enzymes of CNP were also expressed in aT3-1 cells and primary mouse pituitaries. Transcriptional analyses revealed that the proximal 50 bp of the murine Nppc promoter were sufficient for GNRH responsiveness, in an apparent protein kinase C and calcium-dependent manner. Electrophoretic mobility shift assays showed Sp1/Sp3 proteins form major complexes within this region of the Nppc promoter. CNP protein was detectable in rat anterior pituitaries, and electron microscopy detected CNP immunoreactivity in secretory granules of gonadotroph cells. Pharmacological analyses of natriuretic peptide receptor activity clearly showed ANP and CNP are potent activators of cGMP production. However, functional studies failed to reveal a role for CNP in regulating cell proliferation or LH secretion. Surprisingly, CNP potently stimulated the human glycoprotein hormone a-subunit promoter in LbT2 cells but not in aT3-1 cells. Collectively, these findings support a role for CNP as the major natriuretic peptide of the anterior pituitary, and for gonadotroph cells as the major source of CNP expression and site of action.

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Section: Molecular Characterisation Of Gonadotroph Natriuretic Peptidmentioning

confidence: 85%

Molecular characterisation and functional interrogation of a local natriuretic peptide system in rodent pituitaries, αT3-1 and LβT2 gonadotroph cells

Thompson

Chand²,

Jonas

et al. 2009

Journal of Endocrinology

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“…These findings mirror data published by Chinkers and Wilson (21) demonstrating a lack of interaction between the extracellular domains of NPR-A and NPR-B. Furthermore, a dominant-negative mutant of NPR-A as well as an alternative splice variant of NPR-B acting in a dominant-negative manner have previously been used to study ANP and CNP physiology, providing evidence for the use of dominant-negative mutants in cell culture and transgenic animal models as a powerful method to study the specific function of NPRs (19,(22)(23)(24).…”

Section: Discussionmentioning

confidence: 99%

Cardiac hypertrophy in transgenic rats expressing a dominant-negative mutant of the natriuretic peptide receptor B

Langenickel

Buttgereit

Pagel-Langenickel

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

116

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Natriuretic peptides (NP) mediate their effects by activating membrane-bound guanylyl cyclase-coupled receptors A (NPR-A) or B (NPR-B). Whereas the pathophysiological role of NPR-A has been widely studied, only limited knowledge on the cardiovascular function of NPR-B is available. In vitro studies suggest antiproliferative and antihypertrophic actions of the NPR-B ligand C-type NP (CNP). Because of the lack of a specific pharmacological inhibitor, these effects could not clearly be attributed to impaired NPR-B signaling. Recently, gene deletion revealed a predominant role of NPR-B in endochondral ossification and development of female reproductive organs. However, morphological abnormalities and premature death of NPR-B-deficient mice preclude detailed cardiovascular phenotyping. In the present study, a dominant-negative mutant (NPR-B⌬KC) was used to characterize CNP-dependent NPR-B signaling in vitro and in transgenic rats. Here we demonstrate that reduced CNP-but not atrial NP-dependent cGMP response attenuates antihypertrophic potency of CNP in vitro. In transgenic rats, NPR-B⌬KC expression selectively reduced NPR-B but not NPR-A signaling. NPR-B⌬KC transgenic rats display progressive, blood pressure-independent cardiac hypertrophy and elevated heart rate. The hypertrophic phenotype is further enhanced in chronic volume overload-induced congestive heart failure. Thus, this study provides evidence linking NPR-B signaling to the control of cardiac growth.C-type natriuretic peptide ͉ knockdown

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“…Guanylyl cyclase B (GC-B) is another example with two truncated splice forms. It was proposed that these splice forms regulate the function of the full-length subunit and show different tissue distributions (33).…”

Section: Discussionmentioning

confidence: 99%

α1 Soluble Guanylyl Cyclase (sGC) Splice Forms as Potential Regulators of Human sGC Activity

Sharina

Jeleń

Bogatenkova

et al. 2008

Journal of Biological Chemistry

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Soluble guanylyl cyclase (sGC), a key protein in the NO/cGMP signaling pathway, is an obligatory heterodimeric protein composed of one ␣-and one ␤-subunit. The ␣ 1 /␤ 1 sGC heterodimer is the predominant form expressed in various tissues and is regarded as the major isoform mediating NO-dependent effects such as vasodilation. We have identified three new ␣ 1 sGC protein variants generated by alternative splicing. The 363 residue N1-␣ 1 sGC splice variant contains the regulatory domain, but lacks the catalytic domain. The shorter N2-␣ 1 sGC maintains 126 N-terminal residues and gains an additional 17 unique residues. The C-␣ 1 sGC variant lacks 240 N-terminal amino acids, but maintains a part of the regulatory domain and the entire catalytic domain. Q-PCR of N1-␣ 1 , N2-␣ 1 sGC mRNA levels together with RT-PCR analysis for C-␣ 1 sGC demonstrated that the expression of the ␣ 1 sGC splice forms vary in different human tissues indicative of tissue-specific regulation. Functional analysis of the N1-␣ 1 sGC demonstrated that this protein has a dominant-negative effect on the activity of sGC when coexpressed with the ␣ 1 /␤ 1 heterodimer. The C-␣ 1 sGC variant heterodimerizes with the ␤ 1 subunit and produces a fully functional NO-and BAY41-2272-sensitive enzyme. We also found that despite identical susceptibility to inhibition by ODQ, intracellular levels of the 54-kDa C-␣ 1 band did not change in response to ODQ treatments, while the level of 83 kDa ␣ 1 band was significantly affected by ODQ. These studies suggest that modulation of the level and diversity of splice forms may represent novel mechanisms modulating the function of sGC in different human tissues.Since the studies of the late 1970s and early 1980s, which underlined the obligatory role of the endothelium in mediating acetylcholine-induced vasodilatation, nitric oxide (NO) has been recognized as an endogenous nitrovasodilator that mediates the local regulation of basal arterial tone (1-4). Many of the physiological functions of NO in the cardiovascular, neuronal, gastrointestinal, and other systems are mediated through its primary receptor, soluble guanylyl cyclase (sGC).3 The hemecontaining sGC heterodimer converts guanosine triphosphate into the secondary messenger guanosine 3Ј:5Ј-cyclic monophosphate (cGMP). The sGC activity increases more than 200-fold in response to NO (5,6). High concentrations of cGMP produced by activated sGC modulate functions of numerous enzymes, such as cyclic nucleotide phosphodiesterases, cGMP-gated ion channels, and cGMP-dependent protein kinases (PGKs). Recently, the vital importance of sGC for mammalian physiology was directly confirmed by generation of sGC knock-out mice (7-9). The absence of sGC protein resulted in a significant increase in blood pressure, complete loss of NO-dependent aortic relaxation, and inhibition of platelet aggregation in knock-out animals, which died prematurely at the age of 4 weeks because of severe gastrointestinal disorders (7).Four sGC isoforms, products of four genes, have been identified so...

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Regulation of the Guanylyl Cyclase-B Receptor by Alternative Splicing

Cited by 37 publications

References 48 publications

Molecular characterisation and functional interrogation of a local natriuretic peptide system in rodent pituitaries, αT3-1 and LβT2 gonadotroph cells

Molecular characterisation and functional interrogation of a local natriuretic peptide system in rodent pituitaries, αT3-1 and LβT2 gonadotroph cells

Cardiac hypertrophy in transgenic rats expressing a dominant-negative mutant of the natriuretic peptide receptor B

α1 Soluble Guanylyl Cyclase (sGC) Splice Forms as Potential Regulators of Human sGC Activity

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