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Originally published as JHC exPRESS on April 14, 2008.
doi:10.1369/jhc.2008.950733
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Volume 56 (8): 723-731, 2008
Copyright ©The Histochemical Society, Inc.
C9orf10 Protein, a Novel Protein Component of Pur
-containing mRNA-protein Particles (Pur-mRNPs): Characterization of Developmental and Regional Expressions in the Mouse Brain
Research Unit of Biochemistry (YK,KS,SO,KK,KA) and Research Unit of Genome Science (HK), College of Pharmacy, Nihon University, Chiba, Japan; Division of Neuronal Cell Biology, Medical University of Vienna, Center for Brain Research, Vienna, Austria (PM,MK); and Department of Molecular Neurobiology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan (YK)
Correspondence to: Kaijiro Anzai, Research Unit of Biochemistry, College of Pharmacy, Nihon University, 7-7-1, Narashinodai, Funabashi, Chiba 274-8555, Japan. E-mail: kaijiroa{at}pha.nihon-u.ac.jp
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Summary |
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 Top Summary IntroductionMaterials and MethodsResultsDiscussionLiterature Cited |
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Pur
has been implicated in mRNA transport and translation in neurons. We previously reported that Pur is a component of mRNA/protein complexes (Pur-mRNPs) with several other proteins. Among them, we found the C9orf10 (Homo sapiens chromosome 9 open reading frame 10) protein, which was recently characterized as a component of RNA-containing structures. However, C9orf10 itself remains poorly understood. To characterize C9orf10 expression at the protein level, we raised an antibody against C9orf10 and compared the spatial and developmental expressions of this protein and Pur in the mouse brain. C9orf10 was expressed as early as embryo stage 12, whereas Pur was expressed from 5 days after birth. In adults, C9orf10 expression was most prominent in the hippocampus, caudate putamen, cerebral cortex, and cerebellum, unlike the uniform distribution of Pur. C9orf10-positive cells also showed immunoreactivity to Pur. C9orf10 expression was restricted to neurons, judging by the immunoreactivity to neuron-specific nuclear protein or CaM kinase II. These observations suggest an accessory role of C9orf10 for Pur in a limited brain region in addition to other possible functions that have not yet been determined. (J Histochem Cytochem 56:723–731, 2008)
Key Words: Pur • C9orf10 • development • mRNA/protein complexes • brain • mouse
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Introduction |
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TopSummaryIntroductionMaterials and MethodsResultsDiscussionLiterature Cited |
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C9orf10 (Homo sapiens chromosome 9 open reading frame 10) was originally found by the human genome sequence project as an annotated protein, and the gene was mapped to chromosome 9q22.31. Thus far, this protein has been detected in some RNA-containing structures, such as mRNA granules (Bannai et al. 2004
), polyribosomal poly(A)mRNA–mRNA/protein complexes (mRNPs) (Angenstein et al. 2005), and spliceosomes (Rappsilber et al. 2002). However, there is not yet any functional information about this protein.
In a previous study to determine the binding partners of Pur in the neuronal cytoplasm, we isolated polyribosome-containing structures using anti-Pur antibody (Ohashi et al. 2002). Pur has now been found in a multifunctional protein that binds to single-stranded DNA and RNA; it has also been implicated in DNA replication and transcription, cell proliferation (Gallia et al. 2000), mRNA transport (Ohashi et al. 2000,2002; Kanai et al. 2004; Johnson et al. 2006), and translation repression (Gallia et al. 2001).
These biological aspects of Pur may be attributable to various interacting proteins (Johnson et al. 1995; Darbinian et al. 1999; Safak et al. 1999; Tretiakova et al. 1999; Ohashi et al. 2002). We have further examined proteins immunoprecipitated using anti-Pur antibody by proteomic analysis and identified 15 additional proteins (unpublished data), of which a mouse ortholog of human C9orf10 protein was detected. C9orf10 is known to belong to a family of putative transmembrane proteins that includes CXorf17 and BC012177 (Holden and Raymond 2003). CXorf17 has been considered a novel candidate protein for one of the many uncharacterized disorders that map to the human chromosome Xp11.22 (Holden and Raymond 2003), suggesting that C9orf10 may also play a role in neurons. Pur is also known to be indispensable to both postnatal development and the differentiation of certain types of neurons in the mouse brain (Khalili et al. 2003), including synaptogenesis. Thus, it is probable that C9orf10 may, together with Pur, play a crucial role in the development of brain functions. Therefore, it is of great interest to characterize the expression of C9orf10 in the brain in reference to Pur. In this report, we refer to this mouse ortholog as C9orf10 for convenience.
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Materials and Methods |
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TopSummaryIntroductionMaterials and MethodsResultsDiscussionLiterature Cited |
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Sources of Antibodies
Rabbit anti-Pur
antibody was raised by our laboratory and affinity purified as described previously (Ohashi et al. 2002). The affinity purified antibody was used at a concentration of 1:5000 dilutions for Western blot analysis and 1:200 dilutions for immunohistochemistry (IHC). Rabbit anti-C9orf10 antibody was raised in our laboratory against mixed peptides corresponding to amino acids 413-426 (QNSYSNIPHEGKHT; referred to as JB204) and 1062–1075 (TGDPRVPSHSESAL; referred to as JB205) of C9orf10 (NCBI accession number NP_001028440.2), whose sequences have no meaningful homology with other members of this gene family, such as CXorf17 and BC012177, and was affinity purified using the peptides. The affinity purified antibody was used at a concentration of 1:2000 dilutions for Western blot analysis and 1:200 dilutions for IHC.
The other antibodies used were obtained from the following sources. Rabbit anti-S6 ribosomal protein antibody was obtained from Cell Signaling Technologies (Danvers, MA) and used at 1:2000 dilutions for Western blot analysis. Mouse anti-CaM kinase II (CaMKII) antibody, clone 6G9, which reacts with -isoform (Erondu and Kennedy 1985), was purchased from Affinity BioReagents (Golden, CO) and used at 1:100 dilutions for IHC. Mouse anti-neuronal nuclei (NeuN) antibody was purchased from Chemicon International (Temecula, CA) and used at 1:300 dilutions for IHC. Alkaline phosphatase–conjugated anti-rabbit IgG was purchased from Promega (Madison, WI) and used at 1:7500 dilutions for Western blot analysis. Biotinylated anti-rabbit IgG was purchased from Vector Laboratories (Burlingame, CA) and used at 1:200 dilutions for IHC. Alexa Fluor 488–conjugated anti-rabbit IgG and Alexa Fluor 555–conjugated anti-mouse IgG were purchased from Invitrogen (Carlsbad, CA) and used at 1:200 dilutions for IHC.
Animals
Pregnant and adult male mice (ddY strain) were purchased from Japan SLC (Hamamatsu, Japan). The use of animals was approved by the Ethics Review Committee for Animal Experimentation of Nihon University.
Preparation of Tissue Extracts
To obtain mouse brains, mice were killed with an overdose of ether. The brains were quickly removed and immediately washed with ice-cold PBS. To obtain embryo brain samples, pregnant mice were killed with an overdose of ether, and the embryos were quickly removed, washed with ice-cold PBS, and placed on ice in PBS. The embryo brains were quickly removed using forceps under microscope and immediately washed with ice-cold PBS. All the following steps were performed at 0–4C unless otherwise stated. Mouse embryos and adult male brains were homogenized with a Teflon tissue grinder in TKM buffer (20 mM Tris-HCl, pH 7.5, 2 mM MgCl2, 50 mM KCl, 250 mM sucrose) with 1 mM PMSF and 100 µg/ml cycloheximide (CHX), and the homogenate was centrifuged at 10,000 x g for 15 min to yield postmitochondrial supernatant (PMS). The PMS was centrifuged at 130,000 x g for 1 hr, resulting in supernatant (S100) and pellet (P100) fractions. The P100 fraction was resuspended in TKM buffer, and solid KCl was added to a final concentration of 150 mM. The sample was kept on ice for at least 15 min and centrifuged at 10,000 x g for 5 min to remove insoluble materials. The supernatant was used as P100 extracts.
Western Blot Analysis
Western blot analysis was performed as described previously (Ohashi et al. 2000). In brief, samples were electrophoresed and blotted onto a polyvinylidene difluoride (PVDF) membrane. Antibody against protein of interest was incubated with the membrane, and the bound antibody was detected by alkaline phosphatase–conjugated secondary antibody followed by visualization with nitro blue tetrazolium chloride/5-bromo-4-chloro-3'-indolylphosphate p-toluidine salt. The specificity of the anti-C9orf10 antibody was confirmed by preincubation of the antibody with 10- to 100-fold amounts of JB205 peptide overnight at 4C before processing for Western blot analysis.
Immunoprecipitation Analysis
Immunoprecipitation analysis was performed as described previously (Ohashi et al. 2002). In brief, P100 extracts (100 µg of protein in 250 µl) were incubated with 2.5 µg (in 2.5 µl) of the appropriate antibody for 2 hr at 4C in TKM buffer with 100 µg/ml CHX, 1 mM PMSF, and 0.1% Nonidet P-40 (NP40). For P100 extracts subjected to treatment with EDTA, the antibodies were added after the extracts had been treated with or without 25 mM EDTA on ice for 30 min. Subsequently, 40 µl of Dynabeads M-280 sheep anti-rabbit IgG (Invitrogen) was added, and the reaction mixtures were incubated for an additional 4 hr at 4C. The beads were collected with a magnet and washed five times (5 min each time) in 200 µl of TKM buffer containing 0.1% NP40. The beads were washed for an additional 15 min, after which the immune complex was eluted in 40 µl of SDS-containing sample buffer (50 mM Tris-HCl, pH 6.8, 2% SDS, 5% 2-mercaptoethanol, and 10% glycerol). Immunoprecipitated proteins were analyzed by Western blot analysis as described previously (Ohashi et al. 2000).
Fixation of Tissue Samples
To obtain embryo samples, pregnant mice were killed with an overdose of ether, and the embryos were quickly removed. The embryos were fixed with 4% paraformaldehyde (PFA) in PBS. To obtain brain samples, the adult male mice were anesthetized with pentobarbital, and they were perfused transcardially with PBS followed by 4% PFA in PBS. Their brains were dissected and postfixed overnight in 4% PFA in PBS. The embryos and brains were immersed in 30% sucrose in PBS at least overnight. After sinking, they were embedded in Optimal Cutting Temperature Compound (Tissue-Tek; Sakura Finetechnical, Tokyo, Japan), frozen, and sliced at 10 µm by cryostat.
IHC
The sections on glass slides were dried, washed in PBS, and heated at 121C for 3 min in 10 mM sodium citrate buffer (pH 6.0). After the slides were cooled to room temperature, the sections were washed in PBS and incubated in methanol containing 0.3% H2O2 for 30 min to quench endogenous peroxidase activity. The sections were incubated with 2% normal donkey serum in PBS containing 0.3% Tween 20 (blocking solution) at room temperature before processing for IHC. The sections were incubated with the appropriate primary antibody in blocking solution at 4C overnight and were washed with PBS containing 0.1% Tween 20 (PBST). The bound antibodies were detected using the avidin-biotin-peroxidase complex system (Vector Stain Elite ABC Kits; Vector Laboratories) followed by development with DAB (DAB Substrate Kit; Vector Laboratories) in accordance with the manufacturer's instructions.
The specificity of the anti-C9orf10 antibody was confirmed by preincubation of the antibody with a 30-fold amount of JB205 peptide overnight at 4C before processing for IHC. The specificity of the anti- Pur antibody was confirmed by preincubation of the antibody with a 30-fold amount of Pur peptide overnight at 4C before processing for IHC.
Double labeling with anti-C9orf10 and anti-NeuN antibodies or anti-C9orf10 and anti-CaMKII antibodies was accomplished by the simultaneous application of both antibodies. After the sections were incubated with the first antibodies, the sections were incubated with Alexa Fluor 488–conjugated anti-rabbit IgG and Alexa Fluor 555–conjugated anti-mouse IgG for 1 hr at room temperature and washed with PBST. As the negative control, specimens were labeled with the secondary antibodies alone, and none of the control sections showed positive immunoreactivity.
In general, for double labeling performed with antibodies raised against the same species, e.g., rabbit, one-by-one staining with chromogen is well documented (Lan et al. 1995; Xie et al. 2001). However, this method makes it difficult to detect similarities in staining patterns. To study whether or not C9orf10 and Pur are expressed in the same cell, double labeling with anti-C9orf10 and anti-Pur antibodies was conducted under the combination of fluorochrome and chromogen. The first IHC staining for C9orf10 was performed with Alexa Fluor 488–conjugated anti-rabbit IgG and photographed. The slides were soaked in cold PBS overnight to remove the coverslips, followed by washing in PBS and heating at 121C for 3 min in 10 mM sodium citrate buffer (pH 6.0) to inactivate all antibodies bound to the tissue. The second IHC staining for Pur was performed with the avidin-biotin-peroxidase complex system, followed by development with DAB.
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Results |
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TopSummaryIntroductionMaterials and MethodsResultsDiscussionLiterature Cited |
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Characterization of Antibody Against C9orf10 Peptide
To study C9orf10 expression at the protein level in the mouse brain, we raised an antibody against C9orf10 in rabbit using two peptides (JB204 and JB205). Figure 1 shows that Western blot analysis detected one prominent protein band with an apparent molecular mass of 120 kDa. This band disappeared when the antibody was preabsorbed with peptide JB205 but not with peptide JB204 (data not shown), suggesting that the antibody specifically recognized C9orf10 through peptide JB205. This antibody recognized neurons such as Purkinje cells (Figures 2 A and 2B), and immunoreactivity was diminished by preincubation with peptide JB205. These results indicate that the antibody was specific to C9orf10 in Western blot analysis and IHC.
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The specificity of the anti-Pur
antibody was also examined for further experiments. The antibody also recognized Purkinje cells, and the immunoreactivity was diminished by preincubation with Pur peptide (Figures 2C and 2D).
Coimmunoprecipitation of C9orf10 With Pur Using Anti-C9orf10 Antibody
First, we confirmed that C9orf10 is a protein component of Pur-mRNPs using the anti-C9orf10 antibody; C9orf10 had previously been detected by mass spectrometry analysis of proteins pulled down by the anti-Pur antibody, which also contained poly(A)mRNAs (unpublished data). Figure 3
shows that the antibody pulled down Pur, rRNAs, and proteins (S6) and poly(A)mRNAs in addition to C9orf10 (Lane 3), suggesting that polyribosomes containing both proteins were immunoprecipitated. Moreover, no significant reduction was observed in the levels of Pur and poly(A)mRNAs after the P100 extracts were treated with EDTA, but ribosomes (rRNA and S6) were no longer detected (compare Lane 4 with Lane 3). Together, these results suggest that C9orf10 is associated with polyribosomal Pur-mRNPs but not with ribosomes.
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Developmental Expression of C9orf10 in the Mouse Brain
We determined the changes in C9orf10 expression during mouse development. Figure 4 shows that C9orf10 was expressed in the embryonic brain [at 12 days after gestation (E12)] at a level similar to that of neonatal brain (5 days after birth), whereas no Pur
was detected at E12, showing that C9orf10 was expressed at an earlier stage of development than was Pur. C9orf10 expression in the brain increased postnatally and was maintained at an adult level beyond 4 weeks after birth. In contrast to C9orf10, Pur was expressed from 5 days after birth, and the expression was maximal beyond 4 weeks.
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Spatial Expression of C9orf10 in the Mouse Brain
The spatial expression of C9orf10 was analyzed by IHC (Figure 5 ). At 4 weeks after birth, both C9orf10 and Pur
became strongly expressed, but some differences in regional expression were also noted (boxed regions in Figure 5, panels M vs N, O vs P, Q vs R, and S vs T). Higher magnifications of the boxed regions are shown in Figures 5C–5T. Essentially, Pur was detected in all of the brain regions examined, whereas the levels of C9orf10 were much lower in particular regions of the brain, including the superior colliculus (Figure 5M), mid-brain (Figure 5O), pons (Figure 5Q), and the granular layer of the cerebellum (gl in Figure 5S). These results suggest that the regional expression in the brain of C9orf10 is differentially regulated, whereas the C9orf10-expressing region was almost always included in Pur-expressing brain regions.
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Coexpression of C9orf10 With Pur
in Cells in IHCIHC analyses showed that C9orf10 and Pur
proteins were coexpressed in some restricted brain regions (Figure 5). Furthermore, both proteins were coimmunoprecipitated by anti-Pur antibody (data not shown) or anti-C9orf10 antibody (Figure 3). Nevertheless, this does not necessarily mean that C9orf10 and Pur were coexpressed in the same cells. Hence, to unambiguously show the coexpression of the two proteins in cells, we focused on the cerebral cortex in IHC. For simultaneous staining with antibodies against C9orf10 or Pur, we adopted a unique method using fluorescent labeling followed by DAB staining, because both antibodies to be used were raised in rabbits. First, we incubated cells with anti-C9orf10 antibody using Alexa Fluor 488–labeled secondary antibody, followed by heating in citrate buffer, pH 6.0, to inactivate antibodies, and then incubated them again with anti-Pur antibody with biotinylated secondary antibody using the avidin-biotinylated-peroxidase complex (ABC) and DAB. Judging from the immunoreactivity patterns (Figure 6
), anti-C9orf10 antibody (Figures 6A and 6C) and anti-Pur antibody (Figures 6B and 6D) highlighted the same cells. Note also that residual C9orf10 immunoreactivity was abolished by heating, because biotinylated secondary antibody alone did not show immunoreactivity (Figures 6E and 6F). Thus, these results show that C9orf10 and Pur are coexpressed in the same cells in the cerebral cortex.
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Neuron-specific Expression of C9orf10 Protein in the Mouse Brain
The immunoreactivity pattern of C9orf10 suggested that expression was restricted to neuronal cells (Figure 5). As shown in Figure 7 , this was indeed the case, because all of the cells exhibiting C9orf10 immunoreactivity were also detected by NeuN immunoreactivity in the hippocampus (Figures 7A–7C) and cerebral cortex (Figures 7D–7F). Anti-NeuN antibody detects the neuron-specific nuclear protein, NeuN (Mullen et al. 1992
). Furthermore, C9orf10 was also coexpressed with CaMKII, the major postsynaptic density protein (Kennedy et al. 1983), in Purkinje cells (Figures 7G–7I), including the proximal portion of the primary dendrite (indicated by an arrow; inset). These results suggest that C9orf10 could play a role in mRNA metabolism in cooperation with Pur, including transport and translation of particular sets of mRNAs in the neuronal dendrites.
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Discussion |
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TopSummaryIntroductionMaterials and MethodsResultsDiscussionLiterature Cited |
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This study showed that C9orf10 is associated with polyribosomal Pur
-poly(A)mRNPs in a reverse immunoprecipitation analysis using anti-C9orf10 antibody (Figure 3), consistent with others' observations that C9orf10 is present in polyribosome-bound (translated) but not in polyribosome-free (non-translated) fractions from the rat cerebral cortex (Angenstein et al. 2005). We also showed that the expression of C9orf10 was detected as early as E12, which was earlier than Pur and maximal at 4 weeks. At 4 weeks, expression in the brain region was restricted and seemed to be included in the Pur-expressing region (Figure 3). These observations may give some hints on its function. One possible function is that C9orf10 cooperates with Pur in neurons coexpressing the two proteins at a translational level. Another possibility is that C9orf10 may escort Pur-containing mRNA subsets to the cytoplasm after processing nascent mRNAs within nuclei, because the protein is reportedly detected in the human spliceosomes (Rappsilber et al. 2002). The other possibility is that, as has been suggested for Pur (Ohashi et al. 2002; Kanai et al. 2004; Johnson et al. 2006), C9orf10 may also participate in mRNA transport in the cytoplasm. In this respect, C9orf10 is known to associate with an mRNA granule containing SYNCRIP, which carries inositol 1,4,5-triphosphate receptor type 1 mRNA in neuronal dendrites (Bannai et al. 2004). We also previously reported that Pur-mRNPs contain two distinct motor proteins (myosin Va and kinesin 5) in addition to one of the rough endoplasmic reticulum (rER) marker proteins, BiP, and we suggested that vesiculated rERs may be involved in the trafficking of Pur-mRNPs (Ohashi et al. 2002). In this context, C9orf10 could tether Pur-mRNPs to such vesicles that are derived from rER, because C9orf10 has amino acid sequences with the structural features of a transmembrane domain (Holden and Raymond 2003).
In conclusion, our findings suggest that C9orf10, in addition to other possible functions that have not yet been determined, plays a cooperative role with Pur in adult mouse brain neurons. This should encourage precise analyses of Pur-mRNP regulation by C9orf10.
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Acknowledgments |
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This work was supported by a grant from the "Academic Frontiers" Project of the Ministry of Education, Culture, Sports, Science, and Technology of Japan and a Nihon University Research Grant for Assistants and Young Researchers (2005).
We thank Kazusa DNA Research Institute for the KIAA0183 (C9orf10) DNA. We also thank Taeko Takanaka for excellent technical assistance.
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Footnotes |
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Received for publication January 14, 2008; accepted March 24, 2008
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Literature Cited |
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Angenstein F, Evans AM, Ling SC, Settlage RE, Ficarro S, Carrero-Martinez FA, Shabanowitz J, et al. (2005) Proteomic characterization of messenger ribonucleoprotein complexes bound to nontranslated or translated poly(A) mRNAs in the rat cerebral cortex. J Biol Chem 280:6496–6503
Bannai H, Fukatsu K, Mizutani A, Natsume T, Iemura S, Ikegami T, Inoue T, et al. (2004) An RNA-interacting protein, SYNCRIP (heterogeneous nuclear ribonuclear protein Q1/NSAP1) is a component of mRNA granule transported with inositol 1,4,5-trisphosphate receptor type 1 mRNA in neuronal dendrites. J Biol Chem 279:53427–53434
Darbinian N, Gallia GL, Kundu M, Shcherbik N, Tretiakova A, Giordano A, Khalili K (1999) Association of Pur alpha and E2F–1 suppresses transcriptional activity of E2F–1. Oncogene 18:6398–6402[CrossRef][Medline]
Erondu NE, Kennedy MB (1985) Regional distribution of type II Ca2+/calmodulin-dependent protein kinase in rat brain. J Neurosci 5:3270–3277[Abstract]
Gallia GL, Darbinian N, Jaffe N, Khalili K (2001) Single-stranded nucleic acid-binding protein, Pur alpha, interacts with RNA homologous to 18S ribosomal RNA and inhibits translation in vitro. J Cell Biochem 83:355–363[CrossRef][Medline]
Gallia GL, Johnson EM, Khalili K (2000) Puralpha: a multifunctional single-stranded DNA- and RNA-binding protein. Nucleic Acids Res 28:3197–3205
Holden S, Raymond FL (2003) The human gene CXorf17 encodes a member of a novel family of putative transmembrane proteins: cDNA cloning and characterization of CXorf17 and its mouse ortholog orf34. Gene 318:149–161[CrossRef][Medline]
Johnson EM, Chen PL, Krachmarov CP, Barr SM, Kanovsky M, Ma ZW, Lee WH (1995) Association of human Pur alpha with the retinoblastoma protein, Rb, regulates binding to the single-stranded DNA Pur alpha recognition element. J Biol Chem 270:24352–24360
Johnson EM, Kinoshita Y, Weinreb DB, Wortman MJ, Simon R, Khalili K, Winckler B, et al. (2006) Role of Pur alpha in targeting mRNA to sites of translation in hippocampal neuronal dendrites. J Neurosci Res 83:929–943[CrossRef][Medline]
Kanai Y, Dohmae N, Hirokawa N (2004) Kinesin transports RNA: isolation and characterization of an RNA-transporting granule. Neuron 43:513–525[CrossRef][Medline]
Kennedy MB, Bennett MK, Erondu NE (1983) Biochemical and immunochemical evidence that the "major postsynaptic density protein" is a subunit of a calmodulin-dependent protein kinase. Proc Natl Acad Sci USA 80:7357–7361
Khalili K, Del Valle L, Muralidharan V, Gault WJ, Darbinian N, Otte J, Meier E, et al. (2003) Puralpha is essential for postnatal brain development and developmentally coupled cellular proliferation as revealed by genetic inactivation in the mouse. Mol Cell Biol 23:6857–6875
Lan HY, Mu W, Nikolic-Paterson DJ, Atkins RC (1995) A novel, simple, reliable, and sensitive method for multiple immunoenzyme staining: use of microwave oven heating to block antibody crossreactivity and retrieve antigens. J Histochem Cytochem 43:97–102[Abstract]
Mullen RJ, Buck CR, Smith AM (1992) NeuN, a neuronal specific nuclear protein in vertebrates. Development 116:201–211[Abstract]
Ohashi S, Kobayashi S, Omori A, Ohara S, Omae A, Muramatsu T, Li Y, et al. (2000) The single-stranded DNA- and RNA-binding proteins pur alpha and pur beta link BC1 RNA to microtubules through binding to the dendrite-targeting RNA motifs. J Neurochem 75:1781–1790[CrossRef][Medline]
Ohashi S, Koike K, Omori A, Ichinose S, Ohara S, Kobayashi S, Sato TA, et al. (2002) Identification of mRNA/protein (mRNP) complexes containing Puralpha, mStaufen, fragile X protein, and myosin Va and their association with rough endoplasmic reticulum equipped with a kinesin motor. J Biol Chem 277:37804–37810
Rappsilber J, Ryder U, Lamond AI, Mann M (2002) Large-scale proteomic analysis of the human spliceosome. Genome Res 12:1231–1245
Safak M, Gallia GL, Khalili K (1999) Reciprocal interaction between two cellular proteins, Puralpha and YB-1, modulates transcriptional activity of JCVCY in glial cells. Mol Cell Biol 19:2712–2723
Tretiakova A, Steplewski A, Johnson EM, Khalili K, Amini S (1999) Regulation of myelin basic protein gene transcription by Sp1 and Puralpha: evidence for association of Sp1 and Puralpha in brain. J Cell Physiol 181:160–168[CrossRef][Medline]
Xie Y, Nishi S, Iguchi S, Imai N, Sakatsume M, Saito A, Ikegame M, et al. (2001) Expression of osteopontin in gentamicin-induced acute tubular necrosis and its recovery process. Kidney Int 59:959–974[CrossRef][Medline]
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