Human Protein Sam68 Relocalization and Interaction with Poliovirus RNA Polymerase in Infected Cells

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1996 Mar 19

PMID 8637866

Citations 81

Authors

A E McBride

A Schlegel

K Kirkegaard

Affiliations

Soon will be listed here.

Abstract

A HeLa cDNA expression library was screened for human polypeptides that interacted with the poliovirus RNA-dependent RNA polymerase, 3D, using the two-hybrid system in the yeast Saccharomyces cerevisiae. Sam68 (Src-associated in mitosis, 68 kDa) emerged as the human cDNA that, when fused to a transcriptional activation domain, gave the strongest 3D interaction signal with a LexA-3D hybrid protein. 3D polymerase and Sam68 coimmunoprecipitated from infected human cell lysates with antibodies that recognized either protein. Upon poliovirus infection, Sam68 relocalized from the nucleus to the cytoplasm, where poliovirus replication occurs. Sam68 was isolated from infected cell lysates with an antibody that recognizes poliovirus protein 2C, suggesting that it is found on poliovirus-induced membranes upon which viral RNA synthesis occurs. These data, in combination with the known RNA- and protein-binding properties of Sam68, make Sam68 a strong candidate for a host protein with a functional role in poliovirus replication.

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References

Lund P, Simmons J, Hoyt E, DErcole A, Martin F, Van Wyk J . Nucleotide sequence analysis of a cDNA encoding human ubiquitin reveals that ubiquitin is synthesized as a precursor. J Biol Chem. 1985; 260(12):7609-13. View

Bienz K, Egger D, Rasser Y, Bossart W . Intracellular distribution of poliovirus proteins and the induction of virus-specific cytoplasmic structures. Virology. 1983; 131(1):39-48. DOI: 10.1016/0042-6822(83)90531-7. View

Meerovitch K, Svitkin Y, Lee H, Lejbkowicz F, Kenan D, Chan E . La autoantigen enhances and corrects aberrant translation of poliovirus RNA in reticulocyte lysate. J Virol. 1993; 67(7):3798-807. PMC: 237744. DOI: 10.1128/JVI.67.7.3798-3807.1993. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Quadt R, Kao C, Browning K, Hershberger R, Ahlquist P . Characterization of a host protein associated with brome mosaic virus RNA-dependent RNA polymerase. Proc Natl Acad Sci U S A. 1993; 90(4):1498-502. PMC: 45901. DOI: 10.1073/pnas.90.4.1498. View

Morrow C, Gibbons G, Dasgupta A . The host protein required for in vitro replication of poliovirus is a protein kinase that phosphorylates eukaryotic initiation factor-2. Cell. 1985; 40(4):913-21. DOI: 10.1016/0092-8674(85)90351-4. View

Hoffman C, Winston F . A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987; 57(2-3):267-72. DOI: 10.1016/0378-1119(87)90131-4. View

Semler B, Hanecak R, Dorner L, Anderson C, Wimmer E . Poliovirus RNA synthesis in vitro: structural elements and antibody inhibition. Virology. 1983; 126(2):624-35. DOI: 10.1016/s0042-6822(83)80018-x. View

Chien C, Bartel P, Sternglanz R, Fields S . The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci U S A. 1991; 88(21):9578-82. PMC: 52761. DOI: 10.1073/pnas.88.21.9578. View

10.

Evan G, LEWIS G, Ramsay G, Bishop J . Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985; 5(12):3610-6. PMC: 369192. DOI: 10.1128/mcb.5.12.3610-3616.1985. View

11.

Pata J, SCHULTZ S, Kirkegaard K . Functional oligomerization of poliovirus RNA-dependent RNA polymerase. RNA. 1995; 1(5):466-77. PMC: 1482417. View

12.

Andino R, Rieckhof G, Baltimore D . A functional ribonucleoprotein complex forms around the 5' end of poliovirus RNA. Cell. 1990; 63(2):369-80. DOI: 10.1016/0092-8674(90)90170-j. View

13.

Elble R . A simple and efficient procedure for transformation of yeasts. Biotechniques. 1992; 13(1):18-20. View

14.

Bienz K, Egger D, Troxler M, Pasamontes L . Structural organization of poliovirus RNA replication is mediated by viral proteins of the P2 genomic region. J Virol. 1990; 64(3):1156-63. PMC: 249229. DOI: 10.1128/JVI.64.3.1156-1163.1990. View

15.

Bienz K, Egger D, Pasamontes L . Association of polioviral proteins of the P2 genomic region with the viral replication complex and virus-induced membrane synthesis as visualized by electron microscopic immunocytochemistry and autoradiography. Virology. 1987; 160(1):220-6. DOI: 10.1016/0042-6822(87)90063-8. View

16.

Wong G, Muller O, Clark R, Conroy L, Moran M, Polakis P . Molecular cloning and nucleic acid binding properties of the GAP-associated tyrosine phosphoprotein p62. Cell. 1992; 69(3):551-8. DOI: 10.1016/0092-8674(92)90455-l. View

17.

Dewalt P, Johnson V, Lamb J, Semler B . Protein 3CD is the major poliovirus proteinase responsible for cleavage of the P1 capsid precursor. Virology. 1988; 166(1):265-70. DOI: 10.1016/0042-6822(88)90172-9. View

18.

Ruden D, Ma J, Li Y, Wood K, Ptashne M . Generating yeast transcriptional activators containing no yeast protein sequences. Nature. 1991; 350(6315):250-2. DOI: 10.1038/350250a0. View

19.

Jore J, de Geus B, Jackson R, Pouwels P . Poliovirus protein 3CD is the active protease for processing of the precursor protein P1 in vitro. J Gen Virol. 1988; 69 ( Pt 7):1627-36. DOI: 10.1099/0022-1317-69-7-1627. View

20.

Doedens J, Kirkegaard K . Inhibition of cellular protein secretion by poliovirus proteins 2B and 3A. EMBO J. 1995; 14(5):894-907. PMC: 398162. DOI: 10.1002/j.1460-2075.1995.tb07071.x. View