Abstract
The influenza virus nucleoprotein (NP), matrix protein (M1), and ribonucleoproteins (vRNPs) undergo regulated nuclear import and export during infection. Their trafficking was analyzed by using interspecies heterokaryons containing nuclei from infected and uninfected cells. Under normal conditions, it was demonstrated that the vRNPs which were assembled in the nucleus and transported to the cytosol were prevented from reimport into the nucleus. To be import competent, they must first assemble into virions and enter by the endosomal entry pathway. In influenza virus mutant ts51, in which M1 is defective, direct reimport took place but was inhibited by heterologous expression of wild-type M1. These data confirm M1's role as the inhibitor of premature nuclear import and as the main regulator of nuclear transport of vRNPs. In addition to this vRNP shuttling, M1 also shuttled between the nucleus and the cytoplasm in ts51-infected cells. When NP was expressed in the absence of virus infection, it was also found to be a shuttling protein.
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- Adam S. A. The importance of importin. Trends Cell Biol. 1995 May;5(5):189–191. doi: 10.1016/s0962-8924(00)88991-6. [DOI] [PubMed] [Google Scholar]
- Akkina R. K., Chambers T. M., Londo D. R., Nayak D. P. Intracellular localization of the viral polymerase proteins in cells infected with influenza virus and cells expressing PB1 protein from cloned cDNA. J Virol. 1987 Jul;61(7):2217–2224. doi: 10.1128/jvi.61.7.2217-2224.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beaton A. R., Krug R. M. Transcription antitermination during influenza viral template RNA synthesis requires the nucleocapsid protein and the absence of a 5' capped end. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6282–6286. doi: 10.1073/pnas.83.17.6282. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Borer R. A., Lehner C. F., Eppenberger H. M., Nigg E. A. Major nucleolar proteins shuttle between nucleus and cytoplasm. Cell. 1989 Feb 10;56(3):379–390. doi: 10.1016/0092-8674(89)90241-9. [DOI] [PubMed] [Google Scholar]
- Bukrinskaya A. G., Vorkunova N. K., Kornilayeva G. V., Narmanbetova R. A., Vorkunova G. K. Influenza virus uncoating in infected cells and effect of rimantadine. J Gen Virol. 1982 May;60(Pt 1):49–59. doi: 10.1099/0022-1317-60-1-49. [DOI] [PubMed] [Google Scholar]
- Compans R. W., Content J., Duesberg P. H. Structure of the ribonucleoprotein of influenza virus. J Virol. 1972 Oct;10(4):795–800. doi: 10.1128/jvi.10.4.795-800.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davey J., Colman A., Dimmock N. J. Location of influenza virus M, NP and NS1 proteins in microinjected cells. J Gen Virol. 1985 Nov;66(Pt 11):2319–2334. doi: 10.1099/0022-1317-66-11-2319. [DOI] [PubMed] [Google Scholar]
- Davey J., Dimmock N. J., Colman A. Identification of the sequence responsible for the nuclear accumulation of the influenza virus nucleoprotein in Xenopus oocytes. Cell. 1985 Mar;40(3):667–675. doi: 10.1016/0092-8674(85)90215-6. [DOI] [PubMed] [Google Scholar]
- Davidson R. L., Gerald P. S. Improved techniques for the induction of mammalian cell hybridization by polyethylene glycol. Somatic Cell Genet. 1976 Mar;2(2):165–176. doi: 10.1007/BF01542629. [DOI] [PubMed] [Google Scholar]
- Feldherr C. M., Kallenbach E., Schultz N. Movement of a karyophilic protein through the nuclear pores of oocytes. J Cell Biol. 1984 Dec;99(6):2216–2222. doi: 10.1083/jcb.99.6.2216. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fischer U., Meyer S., Teufel M., Heckel C., Lührmann R., Rautmann G. Evidence that HIV-1 Rev directly promotes the nuclear export of unspliced RNA. EMBO J. 1994 Sep 1;13(17):4105–4112. doi: 10.1002/j.1460-2075.1994.tb06728.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldfarb D. S. HIV-1 virology. Simply Marvelous nuclear transport. Curr Biol. 1995 Jun 1;5(6):570–573. doi: 10.1016/s0960-9822(95)00111-4. [DOI] [PubMed] [Google Scholar]
- Goldstein L., Ko C. Distribution of proteins between nucleus and cytoplasm of Amoeba proteus. J Cell Biol. 1981 Mar;88(3):516–525. doi: 10.1083/jcb.88.3.516. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gujuluva C. N., Kundu A., Murti K. G., Nayak D. P. Abortive replication of influenza virus A/WSN/33 in HeLa229 cells: defective viral entry and budding processes. Virology. 1994 Nov 1;204(2):491–505. doi: 10.1006/viro.1994.1563. [DOI] [PubMed] [Google Scholar]
- Hamaguchi M., Maeno K., Nagai Y., Iinuma M., Yoshida T., Matsumoto T. Analysis of nuclear accumulation of influenza nucleoprotein antigen in the presence of p-fluorophenylalanine. Microbiol Immunol. 1980;24(1):51–63. doi: 10.1111/j.1348-0421.1980.tb00561.x. [DOI] [PubMed] [Google Scholar]
- Hamm J., Mattaj I. W. Monomethylated cap structures facilitate RNA export from the nucleus. Cell. 1990 Oct 5;63(1):109–118. doi: 10.1016/0092-8674(90)90292-m. [DOI] [PubMed] [Google Scholar]
- Hay A. J., Wolstenholme A. J., Skehel J. J., Smith M. H. The molecular basis of the specific anti-influenza action of amantadine. EMBO J. 1985 Nov;4(11):3021–3024. doi: 10.1002/j.1460-2075.1985.tb04038.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heinzinger N. K., Bukrinsky M. I., Haggerty S. A., Ragland A. M., Kewalramani V., Lee M. A., Gendelman H. E., Ratner L., Stevenson M., Emerman M. The Vpr protein of human immunodeficiency virus type 1 influences nuclear localization of viral nucleic acids in nondividing host cells. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7311–7315. doi: 10.1073/pnas.91.15.7311. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Helenius A., Marsh M., White J. Inhibition of Semliki forest virus penetration by lysosomotropic weak bases. J Gen Virol. 1982 Jan;58(Pt 1):47–61. doi: 10.1099/0022-1317-58-1-47. [DOI] [PubMed] [Google Scholar]
- Helenius A. Unpacking the incoming influenza virus. Cell. 1992 May 15;69(4):577–578. doi: 10.1016/0092-8674(92)90219-3. [DOI] [PubMed] [Google Scholar]
- Hennekes H., Peter M., Weber K., Nigg E. A. Phosphorylation on protein kinase C sites inhibits nuclear import of lamin B2. J Cell Biol. 1993 Mar;120(6):1293–1304. doi: 10.1083/jcb.120.6.1293. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Izaurralde E., Stepinski J., Darzynkiewicz E., Mattaj I. W. A cap binding protein that may mediate nuclear export of RNA polymerase II-transcribed RNAs. J Cell Biol. 1992 Sep;118(6):1287–1295. doi: 10.1083/jcb.118.6.1287. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones I. M., Reay P. A., Philpott K. L. Nuclear location of all three influenza polymerase proteins and a nuclear signal in polymerase PB2. EMBO J. 1986 Sep;5(9):2371–2376. doi: 10.1002/j.1460-2075.1986.tb04506.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kemler I., Whittaker G., Helenius A. Nuclear import of microinjected influenza virus ribonucleoproteins. Virology. 1994 Aug 1;202(2):1028–1033. doi: 10.1006/viro.1994.1432. [DOI] [PubMed] [Google Scholar]
- Kimura N., Nishida M., Nagata K., Ishihama A., Oda K., Nakada S. Transcription of a recombinant influenza virus RNA in cells that can express the influenza virus RNA polymerase and nucleoprotein genes. J Gen Virol. 1992 Jun;73(Pt 6):1321–1328. doi: 10.1099/0022-1317-73-6-1321. [DOI] [PubMed] [Google Scholar]
- Kistner O., Müller H., Becht H., Scholtissek C. Phosphopeptide fingerprints of nucleoproteins of various influenza A virus strains grown in different host cells. J Gen Virol. 1985 Mar;66(Pt 3):465–472. doi: 10.1099/0022-1317-66-3-465. [DOI] [PubMed] [Google Scholar]
- Kistner O., Müller K., Scholtissek C. Differential phosphorylation of the nucleoprotein of influenza A viruses. J Gen Virol. 1989 Sep;70(Pt 9):2421–2431. doi: 10.1099/0022-1317-70-9-2421. [DOI] [PubMed] [Google Scholar]
- Kobayashi M., Toyoda T., Adyshev D. M., Azuma Y., Ishihama A. Molecular dissection of influenza virus nucleoprotein: deletion mapping of the RNA binding domain. J Virol. 1994 Dec;68(12):8433–8436. doi: 10.1128/jvi.68.12.8433-8436.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kurokawa M., Ochiai H., Nakajima K., Niwayama S. Inhibitory effect of protein kinase C inhibitor on the replication of influenza type A virus. J Gen Virol. 1990 Sep;71(Pt 9):2149–2155. doi: 10.1099/0022-1317-71-9-2149. [DOI] [PubMed] [Google Scholar]
- Laskey R. A., Dingwall C. Nuclear shuttling: the default pathway for nuclear proteins? Cell. 1993 Aug 27;74(4):585–586. doi: 10.1016/0092-8674(93)90505-k. [DOI] [PubMed] [Google Scholar]
- Lewis P. F., Emerman M. Passage through mitosis is required for oncoretroviruses but not for the human immunodeficiency virus. J Virol. 1994 Jan;68(1):510–516. doi: 10.1128/jvi.68.1.510-516.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li R. A., Palese P., Krystal M. Complementation and analysis of an NP mutant of influenza virus. Virus Res. 1989 Feb;12(2):97–111. doi: 10.1016/0168-1702(89)90057-9. [DOI] [PubMed] [Google Scholar]
- Li R. H., Thomas J. O. Identification of a human protein that interacts with nuclear localization signals. J Cell Biol. 1989 Dec;109(6 Pt 1):2623–2632. doi: 10.1083/jcb.109.6.2623. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liljeström P., Garoff H. A new generation of animal cell expression vectors based on the Semliki Forest virus replicon. Biotechnology (N Y) 1991 Dec;9(12):1356–1361. doi: 10.1038/nbt1291-1356. [DOI] [PubMed] [Google Scholar]
- Lin B. C., Lai C. J. The influenza virus nucleoprotein synthesized from cloned DNA in a simian virus 40 vector is detected in the nucleus. J Virol. 1983 Jan;45(1):434–438. doi: 10.1128/jvi.45.1.434-438.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maeda T., Kawasaki K., Ohnishi S. Interaction of influenza virus hemagglutinin with target membrane lipids is a key step in virus-induced hemolysis and fusion at pH 5.2. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4133–4137. doi: 10.1073/pnas.78.7.4133. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maeno K., Yoshii S., Mita K., Hamaguchi M., Yoshida T., Iinuma M., Nagai Y., Matsumoto T. Analysis of the inhibitory effect of canavanine on the replication of influenza RI/5+ virus. I. Inhibition of assembly of RNP. Virology. 1979 Apr 15;94(1):128–137. doi: 10.1016/0042-6822(79)90443-4. [DOI] [PubMed] [Google Scholar]
- Martin K., Helenius A. Nuclear transport of influenza virus ribonucleoproteins: the viral matrix protein (M1) promotes export and inhibits import. Cell. 1991 Oct 4;67(1):117–130. doi: 10.1016/0092-8674(91)90576-k. [DOI] [PubMed] [Google Scholar]
- Martin K., Helenius A. Transport of incoming influenza virus nucleocapsids into the nucleus. J Virol. 1991 Jan;65(1):232–244. doi: 10.1128/jvi.65.1.232-244.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matlin K. S., Reggio H., Helenius A., Simons K. Infectious entry pathway of influenza virus in a canine kidney cell line. J Cell Biol. 1981 Dec;91(3 Pt 1):601–613. doi: 10.1083/jcb.91.3.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mehlin H., Daneholt B., Skoglund U. Translocation of a specific premessenger ribonucleoprotein particle through the nuclear pore studied with electron microscope tomography. Cell. 1992 May 15;69(4):605–613. doi: 10.1016/0092-8674(92)90224-z. [DOI] [PubMed] [Google Scholar]
- Melchior F., Gerace L. Mechanisms of nuclear protein import. Curr Opin Cell Biol. 1995 Jun;7(3):310–318. doi: 10.1016/0955-0674(95)80084-0. [DOI] [PubMed] [Google Scholar]
- Miller J. B. Myoblast diversity in skeletal myogenesis: how much and to what end? Cell. 1992 Apr 3;69(1):1–3. doi: 10.1016/0092-8674(92)90111-o. [DOI] [PubMed] [Google Scholar]
- Murti K. G., Webster R. G., Jones I. M. Localization of RNA polymerases on influenza viral ribonucleoproteins by immunogold labeling. Virology. 1988 Jun;164(2):562–566. doi: 10.1016/0042-6822(88)90574-0. [DOI] [PubMed] [Google Scholar]
- Nassal M., Schaller H. Hepatitis B virus replication. Trends Microbiol. 1993 Sep;1(6):221–228. doi: 10.1016/0966-842x(93)90136-f. [DOI] [PubMed] [Google Scholar]
- Nieto A., de la Luna S., Bárcena J., Portela A., Ortín J. Complex structure of the nuclear translocation signal of influenza virus polymerase PA subunit. J Gen Virol. 1994 Jan;75(Pt 1):29–36. doi: 10.1099/0022-1317-75-1-29. [DOI] [PubMed] [Google Scholar]
- O'Neill R. E., Palese P. NPI-1, the human homolog of SRP-1, interacts with influenza virus nucleoprotein. Virology. 1995 Jan 10;206(1):116–125. doi: 10.1016/s0042-6822(95)80026-3. [DOI] [PubMed] [Google Scholar]
- Ohkuma S., Poole B. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3327–3331. doi: 10.1073/pnas.75.7.3327. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Piñol-Roma S., Dreyfuss G. Shuttling of pre-mRNA binding proteins between nucleus and cytoplasm. Nature. 1992 Feb 20;355(6362):730–732. doi: 10.1038/355730a0. [DOI] [PubMed] [Google Scholar]
- Rees P. J., Dimmock N. J. Kinetics of synthesis of influenza virus ribonucleoprotein structures. J Gen Virol. 1982 Apr;59(Pt 2):403–408. doi: 10.1099/0022-1317-59-2-403. [DOI] [PubMed] [Google Scholar]
- Rey O., Nayak D. P. Nuclear retention of M1 protein in a temperature-sensitive mutant of influenza (A/WSN/33) virus does not affect nuclear export of viral ribonucleoproteins. J Virol. 1992 Oct;66(10):5815–5824. doi: 10.1128/jvi.66.10.5815-5824.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rhoads R. E. Cap recognition and the entry of mRNA into the protein synthesis initiation cycle. Trends Biochem Sci. 1988 Feb;13(2):52–56. doi: 10.1016/0968-0004(88)90028-x. [DOI] [PubMed] [Google Scholar]
- Rihs H. P., Jans D. A., Fan H., Peters R. The rate of nuclear cytoplasmic protein transport is determined by the casein kinase II site flanking the nuclear localization sequence of the SV40 T-antigen. EMBO J. 1991 Mar;10(3):633–639. doi: 10.1002/j.1460-2075.1991.tb07991.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ritchey M. B., Palese P. Identification of the defective genes in three mutant groups of influenza virus. J Virol. 1977 Mar;21(3):1196–1204. doi: 10.1128/jvi.21.3.1196-1204.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruigrok R. W., Calder L. J., Wharton S. A. Electron microscopy of the influenza virus submembranal structure. Virology. 1989 Nov;173(1):311–316. doi: 10.1016/0042-6822(89)90248-1. [DOI] [PubMed] [Google Scholar]
- Ryan K. W., Mackow E. R., Chanock R. M., Lai C. J. Functional expression of influenza A viral nucleoprotein in cells transformed with cloned DNA. Virology. 1986 Oct 15;154(1):144–154. doi: 10.1016/0042-6822(86)90437-x. [DOI] [PubMed] [Google Scholar]
- Schmidt-Zachmann M. S., Dargemont C., Kühn L. C., Nigg E. A. Nuclear export of proteins: the role of nuclear retention. Cell. 1993 Aug 13;74(3):493–504. doi: 10.1016/0092-8674(93)80051-f. [DOI] [PubMed] [Google Scholar]
- Smith G. L., Levin J. Z., Palese P., Moss B. Synthesis and cellular location of the ten influenza polypeptides individually expressed by recombinant vaccinia viruses. Virology. 1987 Oct;160(2):336–345. doi: 10.1016/0042-6822(87)90004-3. [DOI] [PubMed] [Google Scholar]
- Sugiura A., Ueda M., Tobita K., Enomoto C. Further isolation and characterization of temperature-sensitive mutants of influenza virus. Virology. 1975 Jun;65(2):363–373. doi: 10.1016/0042-6822(75)90042-2. [DOI] [PubMed] [Google Scholar]
- Vogel U., Kunerl M., Scholtissek C. Influenza A virus late mRNAs are specifically retained in the nucleus in the presence of a methyltransferase or a protein kinase inhibitor. Virology. 1994 Jan;198(1):227–233. doi: 10.1006/viro.1994.1025. [DOI] [PubMed] [Google Scholar]
- Wakefield L., Brownlee G. G. RNA-binding properties of influenza A virus matrix protein M1. Nucleic Acids Res. 1989 Nov 11;17(21):8569–8580. doi: 10.1093/nar/17.21.8569. [DOI] [PMC free article] [PubMed] [Google Scholar]
- White J., Matlin K., Helenius A. Cell fusion by Semliki Forest, influenza, and vesicular stomatitis viruses. J Cell Biol. 1981 Jun;89(3):674–679. doi: 10.1083/jcb.89.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whittaker G., Bui M., Helenius A. The role of nuclear import and export in influenza virus infection. Trends Cell Biol. 1996 Feb;6(2):67–71. doi: 10.1016/0962-8924(96)81017-8. [DOI] [PubMed] [Google Scholar]
- Whittaker G., Kemler I., Helenius A. Hyperphosphorylation of mutant influenza virus matrix protein, M1, causes its retention in the nucleus. J Virol. 1995 Jan;69(1):439–445. doi: 10.1128/jvi.69.1.439-445.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ye Z. P., Baylor N. W., Wagner R. R. Transcription-inhibition and RNA-binding domains of influenza A virus matrix protein mapped with anti-idiotypic antibodies and synthetic peptides. J Virol. 1989 Sep;63(9):3586–3594. doi: 10.1128/jvi.63.9.3586-3594.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ye Z. P., Pal R., Fox J. W., Wagner R. R. Functional and antigenic domains of the matrix (M1) protein of influenza A virus. J Virol. 1987 Feb;61(2):239–246. doi: 10.1128/jvi.61.2.239-246.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ye Z., Robinson D., Wagner R. R. Nucleus-targeting domain of the matrix protein (M1) of influenza virus. J Virol. 1995 Mar;69(3):1964–1970. doi: 10.1128/jvi.69.3.1964-1970.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]