Phosphorylation of cucumber mosaic virus RNA polymerase 2a protein inhibits formation of replicase complex

The 2a (polymerase) protein of cucumber mosaic virus (CMV) was shown to be phosphorylated both in vivo and in vitro. In vitro assays using 2a protein mutants and tobacco protein kinases showed that the 2a protein has at least three phosphorylation sites, one of which is located within the N‐terminal 126 amino acid region. This region is essential and sufficient for interaction with the CMV 1a protein. When phosphorylated in vitro, the 2a protein N‐terminal region failed to interact with the 1a protein. Since the 1a–2a interaction is essential for the replication of CMV, this suggests that phosphorylation of the N‐terminal region of the 2a protein negatively modulates the interaction in vivo, and may have a regulatory role acting directly in viral infection.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  Viral RNA synthesis by a particulate fraction from cucumber seedlings infected with cucumber mosaic virus. , 1985, Virology.

[3]  R. Symons,et al.  Vira RNA synthesis by a particulate fraction from cucumber seedlings infected with cucumber mosaic virus , 1985 .

[4]  H. Schägger,et al.  Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. , 1987, Analytical biochemistry.

[5]  P. Palukaitis,et al.  Nucleotide sequence and evolutionary relationships of cucumber mosaic virus (CMV) strains: CMV RNA 2. , 1988, The Journal of general virology.

[6]  P Argos,et al.  A sequence motif in many polymerases. , 1988, Nucleic acids research.

[7]  I Sauvaget,et al.  Identification of four conserved motifs among the RNA‐dependent polymerase encoding elements. , 1989, The EMBO journal.

[8]  P. Laybourn,et al.  The transition of RNA polymerase II from initiation to elongation is associated with phosphorylation of the carboxyl-terminal domain of subunit IIa. , 1989, The Journal of biological chemistry.

[9]  V. Blinov,et al.  Two related superfamilies of putative helicases involved in replication, recombination, repair and expression of DNA and RNA genomes. , 1989, Nucleic acids research.

[10]  R. Hayes,et al.  Complete replication of a eukaryotic virus RNA in vitro by a purified RNA-dependent RNA polymerase , 1990, Cell.

[11]  P. Ahlquist,et al.  Use of bromovirus RNA2 hybrids to map cis- and trans-acting functions in a conserved RNA replication gene , 1990, Journal of virology.

[12]  J. Dixon,et al.  Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. , 1991, Analytical biochemistry.

[13]  V. Stollar,et al.  Expression of Sindbis virus nsP1 and methyltransferase activity in Escherichia coli. , 1991, Virology.

[14]  P. Ahlquist,et al.  Brome mosaic virus RNA replication proteins 1a and 2a from a complex in vitro , 1992, Journal of virology.

[15]  W. Rutter,et al.  Interaction cloning: identification of a helix-loop-helix zipper protein that interacts with c-Fos. , 1992, Science.

[16]  R. Schiestl,et al.  Improved method for high efficiency transformation of intact yeast cells. , 1992, Nucleic acids research.

[17]  P. Ahlquist,et al.  Identification of the domains required for direct interaction of the helicase-like and polymerase-like RNA replication proteins of brome mosaic virus , 1992, Journal of virology.

[18]  P. Palukaitis,et al.  Cucumber mosaic virus. , 2018, Advances in virus research.

[19]  S. Fields,et al.  Use of the two-hybrid system to identify the domain of p53 involved in oligomerization. , 1993, Oncogene.

[20]  S. Fields,et al.  Identification of mutations in p53 that affect its binding to SV40 large T antigen by using the yeast two‐hybrid system , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[21]  P. Ahlquist,et al.  Bromovirus RNA replication and transcription require compatibility between the polymerase- and helicase-like viral RNA synthesis proteins , 1993, Journal of virology.

[22]  P. Palukaitis,et al.  The kinetics of infection of zucchini squash by cucumber mosaic virus indicate a function for RNA 1 in virus movement. , 1994, Virology.

[23]  Richard Treisman,et al.  Transcriptional Regulation by Extracellular signals: Mechanisms and Specificity , 1995, Cell.

[24]  T. Hunter,et al.  Transcriptional control by protein phosphorylation: signal transmission from the cell surface to the nucleus , 1995, Current Biology.

[25]  P. Palukaitis,et al.  Viral RNA as a potential target for two independent mechanisms of replicase-mediated resistance against cucumber mosaic virus , 1995, Cell.

[26]  P. Kiberstis,et al.  Multiple serine phosphorylation sites on the 30 kDa TMV cell-to-cell movement protein synthesized in tobacco protoplasts. , 1995, The Plant journal : for cell and molecular biology.

[27]  P. Ahlquist,et al.  Biochemical and genetic analyses of the interaction between the helicase-like and polymerase-like proteins of the brome mosaic virus. , 1995, Virology.

[28]  P. Palukaitis,et al.  Differential effects of satellite RNA on the accumulation of cucumber mosaic virus RNAs and their encoded proteins in tobacco vs zucchini squash with two strains of CMV helper virus. , 1995, Virology.

[29]  Li-li Chen,et al.  A Receptor Kinase-Like Protein Encoded by the Rice Disease Resistance Gene, Xa21 , 1995, Science.

[30]  R. Symons,et al.  An interspecies hybrid RNA virus is significantly more virulent than either parental virus. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[31]  M. Zaitlin,et al.  Cucumber mosaic virus is restricted from entering minor veins in transgenic tobacco exhibiting replicase-mediated resistance. , 1997, Virology.

[32]  C. Kao,et al.  Analysis of the interaction of viral RNA replication proteins by using the yeast two-hybrid assay , 1997, Journal of virology.

[33]  P. Palukaitis,et al.  The plant defense response to cucumber mosaic virus in cowpea is elicited by the viral polymerase gene and affects virus accumulation in single cells , 1997, The EMBO journal.

[34]  S. Dinesh-Kumar,et al.  Signaling in plant-microbe interactions. , 1997, Science.

[35]  Jane Glazebrook,et al.  The Arabidopsis NPR1 Gene That Controls Systemic Acquired Resistance Encodes a Novel Protein Containing Ankyrin Repeats , 1997, Cell.

[36]  C. Kao,et al.  Interactions between the Structural Domains of the RNA Replication Proteins of Plant-Infecting RNA Viruses , 1998, Journal of Virology.

[37]  D. Klessig,et al.  Resistance gene N-mediated de novo synthesis and activation of a tobacco mitogen-activated protein kinase by tobacco mosaic virus infection. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[38]  C. Masuta,et al.  Evolution of a quadripartite hybrid virus by interspecific exchange and recombination between replicase components of two related tripartite RNA viruses. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[39]  Y. Okada,et al.  Phosphorylation and/or Presence of Serine 37 in the Movement Protein of Tomato Mosaic Tobamovirus Is Essential for Intracellular Localization and Stability In Vivo , 1999, Journal of Virology.

[40]  S. Kozlovsky,et al.  Phosphorylation of tobacco mosaic virus movement protein abolishes its translation repressing ability. , 1999, Virology.

[41]  D. Hardie.  PLANT PROTEIN SERINE/THREONINE KINASES: Classification and Functions. , 1999, Annual review of plant physiology and plant molecular biology.

[42]  P. Palukaitis,et al.  The hypersensitive response to cucumber mosaic virus in Chenopodium amaranticolor requires virus movement outside the initially infected cell. , 1999, Virology.

[43]  P. Palukaitis,et al.  Characterisation of genetically modified cucumber mosaic virus expressing histidine-tagged 1a and 2a proteins , 2000, Archives of Virology.

[44]  Erik Andreasson,et al.  Arabidopsis MAP Kinase 4 Negatively Regulates Systemic Acquired Resistance , 2000, Cell.

[45]  V. Citovsky,et al.  Regulation of plasmodesmal transport by phosphorylation of tobacco mosaic virus cell‐to‐cell movement protein , 2000, The EMBO journal.

[46]  S. Blanc,et al.  Evidence for phosphorylation and ubiquitinylation of the turnip yellow mosaic virus RNA-dependent RNA polymerase domain expressed in a baculovirus-insect cell system. , 2000, The Biochemical journal.

[47]  P. Lampe,et al.  Phosphorylation of Connexin43 on Serine368 by Protein Kinase C Regulates Gap Junctional Communication , 2000, The Journal of cell biology.

[48]  M. Zaitlin,et al.  Transgene translatability increases effectiveness of replicase-mediated resistance to cucumber mosaic virus. , 2000, The Journal of general virology.

[49]  Y. Saijo,et al.  Over-expression of a single Ca2+-dependent protein kinase confers both cold and salt/drought tolerance on rice plants. , 2000, The Plant journal : for cell and molecular biology.

[50]  Y. Watanabe,et al.  In vitro phosphorylation of the movement protein of tomato mosaic tobamovirus by a cellular kinase. , 2000, The Journal of general virology.

[51]  G. Martin,et al.  Thr38 and Ser198 are Pto autophosphorylation sites required for the AvrPto–Pto‐mediated hypersensitive response , 2000, The EMBO journal.

[52]  Kwang-Yeol Yang,et al.  Activation of a mitogen-activated protein kinase pathway is involved in disease resistance in tobacco. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[53]  W. J. Lucas,et al.  Phosphorylation of viral movement proteins--regulation of cell-to-cell trafficking. , 2001, Trends in microbiology.

[54]  A. Merits,et al.  Phosphorylation Down-regulates the RNA Binding Function of the Coat Protein of Potato Virus A* , 2001, The Journal of Biological Chemistry.

[55]  D. Klessig,et al.  MAPK cascades in plant defense signaling. , 2001, Trends in plant science.

[56]  D. Tang,et al.  Negative regulation of defense responses in plants by a conserved MAPKK kinase. , 2001, Proceedings of the National Academy of Sciences of the United States of America.