An internal ribosome entry site mediates the initiation of soluble guanylyl cyclase β2 mRNA translation

Vázquez-Padrón, Roberto I.; Pham, Si M.; Mateu, Dania; Khan, Sheik; Aı̈touche, Abdelouahab

doi:10.1111/j.1742-4658.2008.06505.x

Cited by 6 publications

(4 citation statements)

References 34 publications

(36 reference statements)

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“…Each subunit has two isoforms, α1 α2 β1 β2, yet the sGCα1β1 is the most abundant whereas the sGCα2β1 is more predominant in brain tissue [11]. The precise role for β2 subunit is not fully understood and it could have a dominant negative regulatory role [12]. The subunit arrangement for sGCβ1 includes the above mentioned C-terminal CC and GC domains as well as an N-terminal NO-sensing H-NOX/H-NOB and adjacent H-NOXA/H-NOBA/PAS domains.…”

Section: Introductionmentioning

confidence: 99%

Crystal structure of the signaling helix coiled-coil domain of the β1 subunit of the soluble guanylyl cyclase

Beuve

Akker

2010

BMC Struct Biol

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BackgroundThe soluble guanylyl cyclase (sGC) is a heterodimeric enzyme that, upon activation by nitric oxide, stimulates the production of the second messenger cGMP. Each sGC subunit harbor four domains three of which are used for heterodimerization: H-NOXA/H-NOBA domain, coiled-coil domain (CC), and catalytic guanylyl cyclase domain. The CC domain has previously been postulated to be part of a larger CC family termed the signaling helix (S-helix) family. Homodimers of sGC have also been observed but are not functionally active yet are likely transient awaiting their intended heterodimeric partner.ResultsTo investigate the structure of the CC S-helix region, we crystallized and determined the structure of the CC domain of the sGCβ1 subunit comprising residues 348-409. The crystal structure was refined to 2.15 Å resolution.ConclusionsThe CC structure of sGCβ1 revealed a tetrameric arrangement comprised of a dimer of CC dimers. Each monomer is comprised of a long a-helix, a turn near residue P399, and a short second a-helix. The CC structure also offers insights as to how sGC homodimers are not as stable as (functionally) active heterodimers via a possible role for inter-helix salt-bridge formation. The structure also yielded insights into the residues involved in dimerization. In addition, the CC region is also known to harbor a number of congenital and man-made mutations in both membrane and soluble guanylyl cyclases and those function-affecting mutations have been mapped onto the CC structure. This mutant analysis indicated an importance for not only certain dimerization residue positions, but also an important role for other faces of the CC dimer which might perhaps interact with adjacent domains. Our results also extend beyond guanylyl cyclases as the CC structure is, to our knowledge, the first S-helix structure and serves as a model for all S-helix containing family members.

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Section: Introductionmentioning

confidence: 99%

Crystal structure of the signaling helix coiled-coil domain of the β1 subunit of the soluble guanylyl cyclase

Beuve

Akker

2010

BMC Struct Biol

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“…However, the highly folded 5′‐UTR secondary structure might behave as cap‐independent internal ribosome entry site (IRES). The translation efficiency via IRES‐mediated mechanisms found in some viruses [Jackson, 1988; Lourenco et al, 2008] and eukaryotic mRNAs [Hellen and Sarnow, 2001; Vazquez‐Padron et al, 2008] has been known, as the 5′‐UTR sequences contain GC‐rich and complex secondary structures. Except for the likely heat‐activated IRES of that of Drosophila Hsp90 [Ahmed and Duncan, 2004], no IRES structure has so far been described in the 5′‐UTR sequences of rat Hsp90 isoforms.…”

Section: Discussionmentioning

confidence: 99%

Control mechanisms of differential translation of Hsp90 isoforms in 9L rat gliosarcoma cells

Chang

Chao

et al. 2009

J of Cellular Biochemistry

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Although the differential expression of heat shcok proteins, Hsp90alpha and Hsp90beta was extensively studied in many kinds of cells, the post-transcriptional regulation of Hsp90 isoforms remains unclear. In control and GA-treated rat gliosarcoma cells, it has been reported that the translational efficiency of hsp90alpha is higher than hsp90beta. In this study, we present evidences identifying the roles for leaky scanning and 5'-UTR sequence in translational regulation of Hsp90beta. The result of in vitro transcription and translation (IVTT) experiment showed that hsp90alpha exhibited higher translation efficiency than hsp90beta. Sequence analysis revealed that there is an out-of-frame downstream AUG codon in hsp90beta gene. However, elimination of the downstream AUG by site-directly mutagenesis or introducing Kozak context sequence around the initiator AUG of hsp90beta open reading frame increased its translational efficiency, which indicated that leaky scanning might be a possible mechanism regulating hsp90beta. Furthermore, we also constructed a firefly luciferase reporter system to verify the effect of subsequent translation at the downstream out-of-frame AUG codon in 9L and A549 cells. Furthermore, it is believed that 5'-untranslated region (5'-UTR) also plays a significant role in translational control. We showed hsp90beta 5'-UTR gives rise to the reduction of the translation efficiency in IVTT experiment. Additionally, the reductive effect of hsp90beta 5'-UTR was further confirmed by luciferase reporter assay using truncated deletion analyses of 5'-UTR of hsp90beta. Our results support the hypothesis that ribosome leaky scanning mechanism and 5'-UTR sequence acts as negative regulators in hsp90beta mRNA.

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“…Several β2 splice variants have been described in the literature [17, 18, 81, 92]. The human canonical RefSeq gene is comprised of 17 exons (NR_003923) and surprisingly is listed as a non-coding RNA.…”

Section: Regulation Of Sgc Expressionmentioning

confidence: 99%

“…Finally, in a recent report, a new splice variant encoding an ORF with additional amino acids at the N-terminus was described [92]. Interestingly, the same report demonstrated that 5'UTRs of the two alternative β2 sGC transcripts (`canonical' and new splice form) possess an active internal ribosome entry site (IRES).…”

Section: Regulation Of Sgc Expressionmentioning

confidence: 99%

RNA splicing in regulation of nitric oxide receptor soluble guanylyl cyclase

Sharina

Cote²,

Martin

et al. 2011

Nitric Oxide

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Soluble guanylyl cyclase (sGC) is a key protein in the nitric oxide (NO)/-cGMP signaling pathway. sGC activity is involved in a number of important physiological processes including smooth muscle relaxation, neurotransmission and platelet aggregation and adhesion. Regulation of sGC expression and activity emerges as a crucial factor in control of sGC function in normal and pathological conditions. Recently accumulated evidence strongly indicates that the regulation of sGC expression is a complex process modulated on several levels including transcription, post-transcriptional regulation, translation and protein stability. Presently our understanding of mechanisms governing regulation of sGC expression remains very limited and awaits systematic investigation. Among other ways, the expression of sGC subunits is modulated at the levels of mRNA abundance and transcript diversity. In this review we summarize available information on different mechanisms (including transcriptional activation, mRNA stability and alternative splicing) involved in the modulation of mRNA levels of sGC subunits in response to various environmental clues. We also summarize and cross-reference the information on human sGC splice forms available in the literature and in genomic databases. This review highlights the fact that the study of the biological role and regulation of sGC splicing will bring new insights to our understanding of NO/cGMP biology.

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An internal ribosome entry site mediates the initiation of soluble guanylyl cyclase β2 mRNA translation

Cited by 6 publications

References 34 publications

Crystal structure of the signaling helix coiled-coil domain of the β1 subunit of the soluble guanylyl cyclase

Crystal structure of the signaling helix coiled-coil domain of the β1 subunit of the soluble guanylyl cyclase

Control mechanisms of differential translation of Hsp90 isoforms in 9L rat gliosarcoma cells

RNA splicing in regulation of nitric oxide receptor soluble guanylyl cyclase

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