C2PA, a new protein expressed during mouse spermatogenesis

Linares, Jose-Luis; Wendling, Corinne; Tomasetto, Catherine; Rio, Marie‐Christine

doi:10.1016/s0014-5793(00)01942-6

Cited by 8 publications

(12 citation statements)

References 22 publications

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“…Furthermore, because C2PA-RGS3, PDZ-RGS3, and C2PA share eight coding exons, RNA probes originally designed to detect mRNAs encoding mouse PDZ-RGS3 or mouse C2PA may have inadvertently detected transcripts arising from two or three other mRNAs. The probe used to detect transcripts that encode PDZ-RGS3, nucleotides 3-861 of the mouse PDZ-RGS3 cDNA, contained shared nucleotide sequence with other isoforms, and the probes used for northern blot to detect transcripts that encode C2PA (nucleotides 818-2068) and in situ hybridization (nucleotides 818-4712) both overlapped the coding regions for mouse C2PA-RGS3 and PDZ-RGS3 [16,17]. Also, the mRNAs that encode C2PA and PDZ-RGS3 are similar in size making their distinction on northern blot difficult unless specific probes are used.…”

Section: Discussionmentioning

confidence: 99%

“…A direct comparison of the AK004648 nucleotide sequence and the nucleotide sequence that encodes PDZ-RGS3 indicates that AK004648 is a partial cDNA sequence for the PDZ-RGS3 isoform (there are some minor disagreements between the two sequences). A TBLASTN search of the NCBI nonredundant database with the N-terminal 398 amino acids of mouse PDZ-RGS3 revealed that a portion aligned with the coding region for a previously reported open reading frame termed C2PA [17] and the predicted coding region of a human cDNA. Another TBLASTN search with the N-terminal 398 amino acids of mouse PDZ-RGS3 using the NCBI human genome database revealed partial identity with a cluster of likely exons that overlapped the identified exons from which the human C2PA cDNA arose, which just happened to lie 40 kb away from the previously identified RGS3 cluster of exons on chromosome 9.…”

Section: The Identification Of Human Pdz-rgs3mentioning

confidence: 99%

“…As the reported human C2PA coding region only encompassed a portion of mouse C2PA, we used results from BLASTN searches of the NCBI human genome sequences with cDNA sequences that encode human C2PA and mouse C2PA to assemble a predicted C2PA coding region. Like the mouse C2PA [17], the human ortholog has 610 amino acids, a C2 membrane binding domain, an ATP/GTP binding site, and a PDZ domain. The human and mouse proteins share 90% amino acid identity (Fig.…”

Section: The Identification Of Human Pdz-rgs3mentioning

confidence: 99%

“…The 10 upstream exons overlap a previously characterized coding region for a mouse protein termed C2PA [17]. Another RGS3 isoform that uses 17 upstream coding exons along with the same 6 downstream coding exons also exists.…”

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confidence: 98%

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Additional 5′ Exons in the RGS3 Locus Generate Multiple mRNA Transcripts, One of Which Accounts for the Origin of Human PDZ-RGS3

Kehrl¹,

Srikumar²,

Harrison³

et al. 2002

Genomics

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Regulators of G-protein signaling (RGS) proteins can be broadly divided into those that consist predominantly of an RGS domain and those that possess an RGS domain along with additional domains. RGS3 fits into both categories, as both short and longer forms exist. Recently, a novel form of mouse RGS3 that possesses a PDZ domain was identified. Here we show that the human PDZ-RGS3 isoform arises from 10 upstream exons along with 6 exons from the previously characterized RGS3. We found that 47,000 nucleotides span the last of the 10 upstream exons and the first exon used from the original cluster of RGS3 exons. These 10 upstream exons encode 398 amino acids, which show strong conservation with those from mouse PDZ-RGS3. In addition, another isoform exists that uses 17 upstream exons, 9 of which overlap with those in PDZ-RGS3, along with the same 6 downstream exons used in PDZ-RGS3. Finally, a short form of human RGS3 arises from an unrecognized RGS3 exon that encodes an amino-terminal 140 amino acids. For each RGS3 isoform, RT-PCR detected specific mRNA transcripts and immunoblot analysis identified specific bands for RGS3 and PDZ-RGS3. RGS3 provides an example of the complex origins of the coding regions of mammalian proteins.Key Words: RGS, RNA splicing, G-protein, genomic amino-terminal region that shares some similarity with a domain in synapsin [5]. Northern blot analysis revealed prominent 1.8-and 3.5-kb RGS3 mRNA transcripts, but also two lessprominent transcripts of 4.2 and 4.5 kb [5]. The 1.8-and 3.5-kb mRNA transcripts presumably encoded short and long versions of RGS3, but the larger transcripts had no protein-coding region assigned to them. An expression vector for a short form of RGS3 (RGS3T or RGS3CT) was created by arbitrarily choosing an internal ATG [5][6][7]. Both forms of RGS3 act as GAPs for G i␣ and G q␣ , but not G s␣ or G 12␣ [8]. Expression of either form of RGS3 inhibits signaling through a variety of GPCRs that use G i␣ and G q␣ to tranduce signals [5][6][7][8][9]. The shorter version of RGS3 in part localizes in the nucleus and its expression promotes apoptosis of Chinese hamster ovary (CHO) cells [10]. The introduction of RGS3 into B-cell lines inhibited CXCR1-and CXCR4-directed cell migration [11,12]. Perhaps accounting for the efficacy of RGS3 as an inhibitor of chemotaxis, it, in contrast to several other RGS proteins, also inhibits signaling by free G ␤1␥2 subunits [13]. Interference with G ␤␥ signaling blocks chemokine-directed migration [14,15]. 861Article doi:10.1006/geno.2002.6773, available online at http://www.idealibrary.com on IDEAL A partial cDNA for the mouse long form of RGS3 and for a short version of RGS3 with a unique N-terminal 21-amino-acid extension [12] have been identified, as well as a cDNA for the purported full-length version of the long form of mouse RGS3 (GenBank acc. no. AK004648). A partial cDNA clone for another isoform of mouse RGS3, which possesses a 398-amino-acid N-terminal extension containing a PDZ domain, was isolated by interaction cloning using t...

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Section: Discussionmentioning

confidence: 99%

Section: The Identification Of Human Pdz-rgs3mentioning

confidence: 99%

Section: The Identification Of Human Pdz-rgs3mentioning

confidence: 99%

mentioning

confidence: 98%

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Additional 5′ Exons in the RGS3 Locus Generate Multiple mRNA Transcripts, One of Which Accounts for the Origin of Human PDZ-RGS3

Kehrl¹,

Srikumar²,

Harrison³

et al. 2002

Genomics

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“…SRB-RGS bears a PDZ domain that is a multi-functional proteinprotein interaction module that plays important roles in organizing signal transduction complexes, clustering membrane proteins, and maintaining cell polarity at the NH 2 -terminal, as well. 16,17) rERaC/D interacted in vitro with partial SRB-RGS1-495 amino acid (a.a.). SRB-RGS suppressed the transcriptional activities of ERa and the other promoters.…”

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confidence: 99%

Characterization and Identification of a Steroid Receptor-Binding Protein, SRB-RGS

Ikeda

Inoue

Muramatsu³

et al. 2007

Biological & Pharmaceutical Bulletin

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The nuclear hormone receptors are a superfamily of ligand-inducible transcription factors. They include the receptor proteins for steroids, thyroid hormone, vitamin D, and retinoids. Steroid hormone receptors are structurally related and composed of six major functional domains. Domain A/B in the NH 2 -terminal region of the protein is the constitutive activation 1 (AF-1). Domain C, the DNA-binding domain, is arranged in two zinc-stabilized DNA-binding finger motifs. Region D contains a nuclear localization signal. The ligandbinding and ligand-dependent transcriptional activation function 2 (AF-2) is located in domain E/F of the COOH-terminal region of the protein. [1][2][3] For the activation of transcription, steroid hormone receptors recruit coactivators such as SRC-1/p160, p300/CBP, ARA70, Tip60, and RIP140, through binding to AF-1 and AF-2 of the receptors. This kind of coactivator recruits histone acetyltransferase (HAT) or bear HAT activity on itself. Recruitment of HAT allows the local decondensation of chromatin. In the next step, the DRIP/TRAP complex binds to the AF-2 of the receptor and mediates transcriptional activation. This kind of coactivator does not recruit HAT. Thus, these coactivators facilitate the transcription process. [4][5][6][7][8][9] The thyroid hormone receptor (TR) and retinoid X receptor (RXR) in the absence of ligand bind to the corepressors, such as N-CoR and SMRT, respectively, suppressing transcriptional activity. The corepressors recruit histone deacetylase (HDAC).10-13) The chromatin-modifying complexes, ATP-dependent remodeling complexes and HAT or HDAC complexes regulate chromatin structure and transcription so that they might be coordinated to create a DNA template that is accessible to the general transcription apparatus. 14)We employed a yeast two-hybrid system for cloning the cDNA of the protein that interacted with domains C and D of the rat estrogen receptora (rERaC/D) from the rat ovary cDNA library and cloned the cDNA of a novel steroid receptor-binding (SRB) protein bearing the regulator of the G-protein signaling (RGS) domain 15) at the COOH-terminal, designated as SRB-RGS. RGS proteins are GTPase accelerating proteins, which interact with the Ga-subunits that are linked to seven transmembrane G-protein coupled receptor. SRB-RGS bears a PDZ domain that is a multi-functional proteinprotein interaction module that plays important roles in organizing signal transduction complexes, clustering membrane proteins, and maintaining cell polarity at the NH 2 -terminal, as well. 16,17) rERaC/D interacted in vitro with partial SRB-RGS1-495 amino acid (a.a.). SRB-RGS suppressed the transcriptional activities of ERa and the other promoters. 18)We could show that the full-length SRB-RGS interacted with either the full-length ERa or ERb by a coimmunoprecipitation assay. The full-length SRB-RGS and full length ERa interacted in COS-7 cell by a mammalian two-hybrid system. The interaction between intrinsic SRB-RGS and ERs in the nuclear ER extract from the rat uteri was suggested b...

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C2PA is a nuclear protein implicated in the heat shock response

Hirabayashi

Ohno

Iida

et al. 2002

J of Cellular Biochemistry

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C2PA is a protein of unknown function that is expressed in spermatocytes. PDZ-RGS3 is a signaling molecule whose PDZ domain binds Ephrin-B2 and mediates reverse signaling of this protein. C2PA and PDZ-RGS3 have identical PDZ domains. To explore the function of C2PA, we compared it with PDZ-RGS3 with respect to tissue distribution, subcellular localization, and biochemistry. C2PA is expressed only in testis, whereas PDZ-RGS3 is expressed in various tissues including brain, heart, lung, liver, spleen, kidney, small intestine, skeletal muscles, and testis. These proteins also differ in their subcellular distribution, in that PDZ-RGS3 is cytosolic while C2PA is exclusively nuclear. C2PA is distributed diffusely in the nucleus and forms a few foci at 37 degrees C. However, when cells are exposed to 42 degrees C, the number of C2PA foci is increased. These heat shock-induced foci colocalize with CREB-binding protein and heat shock factor-1. In contrast, the distribution of PDZ-RGS3 does not change during heat stress. When overexpressed, C2PA induces heat shock response element (HSE)-dependent gene transcription, whereas PDZ-RGS3 does not. These data suggest that the function of C2PA is distinct from that of PDZ-RGS3, and that C2PA may be involved in the heat shock response in testis.

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C2PA, a new protein expressed during mouse spermatogenesis

Cited by 8 publications

References 22 publications

Additional 5′ Exons in the RGS3 Locus Generate Multiple mRNA Transcripts, One of Which Accounts for the Origin of Human PDZ-RGS3

Additional 5′ Exons in the RGS3 Locus Generate Multiple mRNA Transcripts, One of Which Accounts for the Origin of Human PDZ-RGS3

Characterization and Identification of a Steroid Receptor-Binding Protein, SRB-RGS

C2PA is a nuclear protein implicated in the heat shock response

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