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Expression of PACT and EIF2C2, Implicated in RNAi and MicroRNA Pathways, in Various Human Cell Lines  

Lee, Yong-Sun (Department of Biochemistry and Molecular Genetics, University of Virginia, Institute of Molecular Biology and Genetics, School of Biological Sciences, Seoul National University)
Jeon, Yesu (Department of Biochemistry and Molecular Genetics, University of Virginia, Institute of Molecular Biology and Genetics, School of Biological Sciences, Seoul National University)
Park, Jong-Hoon (Department of Biochemistry and Molecular Genetics, University of Virginia, Institute of Molecular Biology and Genetics, School of Biological Sciences, Seoul National University)
Hwang, Deog-Su (Institute of Molecular Biology and Genetics, School of Biological Sciences, Seoul National University)
Dutta, Anindya (Department of Biochemistry and Molecular Genetics, University of Virginia)
Publication Information
Animal cells and systems / v.8, no.3, 2004 , pp. 213-220 More about this Journal
Abstract
MicroRNA and siRNA (small interfering RNA), representative members of small RNA, exert their effects on target gene expression through association with protein complexes called miRNP (microRNA associated ribonucleoproteins) and RISC (RNA induced silencing complex), respectively. Although the protein complexes are yet to be fully characterized, human EIF2C2 protein has been identified as a component of both miRNP and RISC. In this report, we raised antiserum against EIF2C2 in order to begin understanding the protein complexes. An immunoblot result indicates that EIF2C2 protein is ubiquitously expressed in a variety of cell lines from human and mouse. EIF2C2 protein exists in both cellular compartments, as indicated by an immunoblot assay with a nuclear extract and a cytosolic fraction (S100 fraction) from HeLa S3 lysate. Depletion of EIF2C1 or EIF2C2 protein resulted in a decrease of microRNA, suggesting a possible role of these proteins in microRNA stability or biogenesis. We also prepared antiserum against dsRNA binding protein PACT, whose homologs in C. elegans and Drosophila are known to have a role in the RNAi (RNA interference) pathway. The expression of PACT protein was also observed in a wide range of cell lines.
Keywords
MicroRNA; RNA interference; EIF2C2; PACT; Antibody;
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1 Hannon GJ (2002) RNA interference. Nature 418: 244-251   DOI   ScienceOn
2 Hutvagner G, McLachlan J, Pasquinelli AE, Balint E, Tuschl T, and Zamore PD (2001) A cellular function for the RNAinterference enzyme dicer in the maturation of the let-7 small temporal RNA. Science 293: 834-838   DOI   ScienceOn
3 Carmell MA, Xuan Z, Zhang MQ and Hannon GJ (2002) The Argonaute family: tentacles that reach into RNAi, developmental control, stem cell maintenance, and tumorigenesis. Genes Dev 16: 2733-2742   DOI   ScienceOn
4 Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, and Mello CC (1998) Potent and specific genetic interference by double-stmnded RNA in Caenorhabditis elegans. Nature 391: 806-811   DOI   ScienceOn
5 Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116: 281-297   DOI   ScienceOn
6 Billy E, Brondani V, Zhang H, Muller U and Filipowicz W (2001) Specific interference with gene expression induced by long, double-stranded RNA in mouse embryonal teratocarcinoma cell lines. Proc Natl Acad Sci USA 98: 14428-14433   DOI   ScienceOn
7 Hammond SM, Boettcher S, Caudy AA, Kobayashi R, and Hannon GJ (2001) Argonaute2, a link between genetic and biochemical analyses of RNAi. Science 293: 1146-1150   DOI   ScienceOn
8 Hammond SM, Bernstein E, Beach D, and Hannon GJ (2000) An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells. Nature 404: 293-296   DOI   ScienceOn
9 Gupta V and Patel RC (2002) Proapoptotic protein PACT is expressed at high levels in colonic epithelial cells in mice. Am J Physiol Gastrointest Liver Physiol 283: G801-808   DOI   ScienceOn
10 Fukagawa T, Nogami M, Yoshikawa M, Ikeno M, Okazaki T, Takami Y, Nakayama T, and Oshimura M (2004) Dicer is essential for formation of the heterochromatin structure in vertebrate cells. Nat Cell Biol 6: 784-791   DOI   ScienceOn
11 Saxena S, Jonsson ZO, and Dutta A (2003) Small RNAs with imperfect match to endogenous mRNA repress translation. Implications for off-target activity of small inhibitory RNA in mammalian cells. J Biol Chem 278: 44312-44319   DOI   ScienceOn
12 Ekwall K (2004) The RITS complex-A direct link between small RNA and heterochromatin. Mol Cell 13: 304-305   DOI   ScienceOn
13 Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, and Tuschl T (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411: 494-498   DOI   ScienceOn
14 Dykxhoorn DM, Novina CD, and Sharp PA (2003) Killing the messenger: short RNAs that silence gene expression. Nat Rev Mol Cell Biol 4: 457-467   DOI   ScienceOn
15 Doi N, Zenno S, Ueda R, Ohki-Hamazaki H, Ui-Tei K, and Saigo K (2003) Shortinterfering-RNA-mediated gene silencing in mammalian cells requires Dicer and elF2C translation initiation factors. Curr Biol 13: 41-46   DOI   ScienceOn
16 Doench JG, Petersen CP, and Sharp PA (2003) siRNAs can function as miRNAs. Genes Dev 17: 438-442   DOI   ScienceOn
17 Cerutti L, Mian N, and Bateman A (2000) Domains in gene silencing and cell differentiation proteins: the novel PAZ domain and redefinition of the Piwi domain. Trends Biochem Sci 25: 481-482   DOI   ScienceOn
18 Song JJ, Liu J, Tolia NH, Schneiderman J, Smith SK, Martienssen RA, Hannon GJ, and Joshua-Tor L (2003) The crystal structure of the Argonaute2 PAZ domain reveals an RNA binding motif in RNAi effector complexes. Nat Struct Biol 10: 1026-1032   DOI   ScienceOn
19 Schwarz DS and Zamore PD (2002) Why do miRNAs live in the miRNP? Genes Dev 16: 1025-1031   DOI   ScienceOn
20 Dignam JD, Lebovitz RM, and Roeder RG (1983b) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acid Res 11: 1475-1489   DOI   ScienceOn
21 Lee RC, Feinbaum RL, and Ambros V (1993) The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 75: 843-854   DOI   ScienceOn
22 Sambrook J, Fritsch EF, and Maniatis T (1989) Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor
23 Sasaki T, Shiohama A, Minoshima S, and Shimizu N (2003) Identification of eight members of the Argonaute family in the human genome small star, filled. Genomics 82: 323-330   DOI   ScienceOn
24 Reinhart BJ, Slack FJ, Basson M, Pasquinelli AE, Bettinger JC, Rougvie AE, Horvitz HR, and Ruvkun G (2000) The 21nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans. Nature 403: 901-906   DOI   ScienceOn
25 Patel RC and Sen GC (1998) PACT, a protein activator of the interferon-induced protein kinase, PKR. EMBO J 17: 4379-4390   DOI   ScienceOn
26 Okamura K, Ishizuka A, Siomi H, and Siomi MC (2004) Distinct roles for Argonaute proteins in small RNA-directed RNA cleavage pathways. Genes Dev 18: 1655-1666   DOI   ScienceOn
27 Mourelatos Z, Dostie J, Paushkin S, Sharma A, Charroux B, Abel L, Rappsilber J, Mann M, and Dreyfuss G (2002) miRNPs: a novel class of ribonucleoproteins containing numerous microRNAs. Genes Dev 16: 720-728   DOI   ScienceOn
28 Martinez J, Patkaniowska A, Urlaub H, Luhrmann Rand Tuschl T (2002) Singlestranded antisense siRNAs guide target RNA cleavage in RNAi. Cell 110: 563-574   DOI   ScienceOn
29 Lingel A, Simon B, Izaurralde E, and Sattler M (2003) Structure and nucleic-acid binding of the Drosophila Argonaute 2 PAZ domain. Nature 426: 465-469   DOI   ScienceOn
30 Liu Q, Rand TA, Kalidas S, Du F, Kim HE, Smith DP, and Wang X (2003) R2D2, a bridge between the initiation and effector steps of the Drosophila RNAi pathway. Science 301: 1921-1925   DOI   ScienceOn
31 Tang G, Reinhart BJ, Bartel DP, and Zamore PD (2003) A biochemical framework for RNA silencing in plants. Genes Dev 17: 49-63   DOI   ScienceOn
32 Stark GR, Kerr IM, Williams BR, Silverman RH, and Schreiber RD (1998) How cells respond to interferons. Annu Rev Biochem 67: 227-264   DOI   ScienceOn
33 Tabara H, Sarkissian M, Kelly WG, Fleenor J, Grishok A, Timmons L, Fire A, and Mello CC (1999) The rde-1 gene, RNA interference, and transposon silencing in C. elegans. Cell 99: 123-132   DOI   ScienceOn
34 Tabara H, Yigit E, Siomi H, and Mello CC (2002) The dsRNA binding protein RDE-4 interacts with RDE-1, DCR-1, and a DExH-box helicase to direct RNAi in C. elegans. Cell 109: 861-871   DOI   ScienceOn
35 Yang S, Tutton S, Pierce E, and Yoon K (2001) Specific doublestranded RNA interference in undifferentiated mouse embryonic stem cells. Mol Cell Biol 21: 7807-7816   DOI   ScienceOn
36 Zeng Y and Cullen BR (2002) RNA interference in human cells is restricted to the cytoplasm. RNA 8: 855-860   DOI
37 Zeng Y, Wagner EJ, and Cullen BR (2002) Both natural and designed micro RNAs can inhibit the expression of cognate mRNAs when expressed in human cells. Mol Cell 9: 1327-1333   DOI   ScienceOn
38 Hutvagner G and Zamore PD (2002) A microRNA in a multipleturnover RNAi enzyme complex. Science 297: 2056-2060   DOI   ScienceOn
39 Ishizuka A, Siomi MC, and Siomi H (2002) A Drosophila fragile X protein interacts with components of RNAi and ribosomal proteins. Genes Dev 16: 2497-2508   DOI   ScienceOn