Exploración de la interacción entre la región 5´UTR del Sugarcane Mosaic Virus (SCMV) y proteínas del hospedero maíz

Exploración de la interacción entre la región 5´UTR del Sugarcane Mosaic Virus (SCMV) y proteínas del hospedero maíz

Main Article Content

Giovanni Chaves-Bedoya
Abstract

 

Antecedentes: El virus del mosaico de la caña de azúcar, SCMV (Potyviridae) es un miembro de la gran familia de virus de ARN de cadena positiva, sin una estructura CAP en su extremo 5´no traducido (5`UTR), pero con una proteína viral unida al genoma (VPg) y una cola poli A en su extremo 3´UTR. Se ha sugerido que proteínas del hospedero hacen un puente entre las regiones no traducidas virales 5´y 3´ para potenciar la traducción viral. Dado que las regiones no traducidas presentes en los genomas virales contienen elementos involucrados en la regulación de su ciclo replicativo, es importante analizar la interacción entre estas regiones y las proteínas virales o del hospedero para inferir su función. Objetivo: Determinar si existen proteínas del hospedero maíz que pudieran estar interactuando con la región 5´UTR de SCMV. Metodología: La región 5´UTR de SCMV se amplificó por PCR incluyendo la secuencia del promotor T7 en la secuencia del oligo 5´ UTR y se generaron sus transcritos marcados radiactivamente. Los transcritos marcados fueron entrecruzados con extractos proteicos de hojas de maíz sanas e infectadas. Resultados: Los resultados sugieren la presencia de una proteína putativa de interacción del hospedante maíz de aproximadamente 53kDa, con la región 5´UTR de SCMV. Conclusión: Los ensayos de entrecruzamiento, especificidad y estabilidad de los complejos ARN-proteína sugieren que en el hospedante maíz existe al menos una proteína que interactúa con la región 5´ UTR de SCMV.

Palabras Clave: Complejo ARN-proteína, entrecruzamiento UV, fracción proteica, Potyvirus, VPg.

 

Abstract


Background: Sugarcane mosaic virus is a member of the great family of positive sense RNA viruses, without a CAP structure in its 5´UTR end, but with a viral protein attached to the genome (VPg) and a poli A tail in its 3´UTR end. It has been suggested that some host proteins make a bridge between the untranslated 5´and 3´ regions (UTRs) to enhance the canonical and/or non-canonical virus translation. Since the UTR regions present in the viral genomes have some elements involved in the regulation of their replicative cycle, it is important to analyze the interaction that may occur between these regions and the viral or host proteins to infer regarding its function. Objective: To identify proteins of the host maize that could be interacting with the 5´UTR region of SCMV. Methodology: 5´UTR region of SCMV was amplified by PCR including the sequence of the T7 promoter in the 5´oligo sequence. The transcripts were radioactively labeled. Labeled transcripts were crosslinked with proteins extracts from healthy and infected maize leaves. Results: The results suggest the presence of a 53 kDa putative protein interacting with the 5 ´UTR region of SCMV. Conclusion: Crosslinking essays, specificity and the stability of the RNA-protein complex suggest that in maize host there is at least one protein that interacts with the 5´UTR region of SCMV.

Keywords: RNA-Protein complex, UV crosslinking, protein fraction, Potyvirus, VPg.

 

Resumo


Antecedentes: O vírus do mosaico da cana de açúcar (SCMV) da Família Potyviridae é um membro da grande família de vírus de ARN de cadeia positiva, sem uma estrutura CAP em seu extremo 5´ não traduzido (5`UTR), mas com uma proteína viral unida ao genoma (VPg) e uma cola poli A em seu extremo 3´UTR. Tem-se sugerido que proteínas do hospedeiro fazem uma ponte entre as regiões não traduzidas virales 5´e 3´ para potenciar a tradução viral. Devido que as regiões não traduzidas presentes nos genomas virais contém elementos involucrados na regulação de seu ciclo replicativo, é importante analisar a interação entre estas regiões e as proteínas virales O do hospedeiro para inferir sua função. Objetivo: Determinar se existem proteínas hospedeiras milho que puderam estar interatuando com a região 5´UTR de SCMV. Metodologia: A região 5´UTR de SCMV se amplificou por PCR incluindo a sequência do promotor T7 na sequência do oligo 5´ UTR e se geraram seus transcritos marcados radiativamente. Os transcritos marcados foram entrecruzados com extratos proteicos de folhas de milho saudáveis e infetadas. Resultados: Os resultados sugerem a presença de uma proteína putativa de interação do hospedeiro milho de aproximadamente 53 kDa, com a região 5´UTR de SCMV. Conclusão: Os ensaios de entrecruzamento, especificidade e estabilidade dos complexos ARN proteía sugerem que no hospedeiro milho existe pelo menos uma proteína que interatua com a região 5´ UTR de SCMV.

Palavras-chave: complexo ARN-proteína, entrecruzamento UV, fracção proteica, Potyvirus, VPg.

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Article Details

Author Biography (SEE)

Giovanni Chaves-Bedoya, Universidad Francisco de Paula Santander

Doctorado en Biotecnología Vegetal. Grupo de Investigación PLANTAE, Facultad de Ciencias Básicas.
References

D.D. Shukla, M. Tosic, J.M. Jilka, R. Ford, W. Toler, and A. Langham, “Taxonomy of potyvirus infecting maize, sorhum, and sugarcane in Australia and the united States as determined by reactivities of polyclonal antibodies directed towards virusspecific N termini of coat proteins”, Phytopatology, vol. 79, pp. 223-229, 1989.

P. Ahlquist, A.O. Noueiry, W.M. Lee, D.B. Kushner and B.T. Dye, “Host factors in positive-strand RNA virus genome replication”, J Virol, vol. 77, pp. 8181-6, Aug 2003.

S.U. Huh and K.H. Paek, “Plant RNA binding proteins for control of RNA virus infection,” Front Physiol, vol. 4, pp. 397, 2013.

J.C. Carrington, K.D. Kasschau, S.K. Mahajan and M.C. Schaad, “Cell-toCell and Long-Distance Transport of Viruses in Plants,” Plant Cell, vol. 8, pp. 1669-1681, Oct 1996.

J.L. Riechmann, M.T. Cervera and J.A. Garcia, “Processing of the plum pox virus polyprotein at the P3-6K1 junction is not required for virus viability,” J Gen Virol, vol. 76 ( Pt 4), pp. 951-6, Apr 1995.

S. Urcuqui-Inchima, J. Walter, G. Drugeon, S. German-Retana, A. L. Haenni, T. Candresse, et al, “Potyvirus helper component-proteinase selfinteraction in the yeast two-hybrid system and delineation of the interaction domain involved,” Virology, vol. 258, pp. 95-9, May 25 1999.

B.C. Kang, I. Yeam, J.D. Frantz, J.F. Murphy and M.M. Jahn, “The pvr1 locus in Capsicum encodes a translation initiation factor eIF4E that interacts with Tobacco etch virus VPg,” Plant J, vol. 42, pp. 392 405, May 2005.

K. Thivierge, V. Nicaise, P.J. Dufresne, S. Cotton, J.F. Laliberte, O.Le Gall, et al, “Plant virus RNAs. Coordinated recruitment of conserved host functions by (+) ssRNA viruses during early infection events,” Plant Physiol, vol. 138, pp. 1822-7, Aug 2005.

P.D. Nagy and J. Pogany, “The dependence of viral RNA replication on co-opted host factors,” Nat Rev Microbiol, vol. 10, pp. 137-49, Dec 19 2011.

D. Guo, M.L. Rajamaki, M. Saarma and J.P. Valkonen, “Towards a protein interaction map of potyviruses: protein interaction matrixes of two potyviruses based on the yeast two-hybrid system,” J Gen Virol, vol. 82, pp. 935-9, Apr 2001.

C. Robaglia and C. Caranta, “Translation initiation factors: a weak link in plant RNA virus infection,” Trends Plant Sci, vol. 11, pp. 40-5, Jan 2006.

J. Diaz-Pendon, V. Truniger, C. Nieto, J. Garcia-Mas, A. Bendahmane and M. A. Aranda, “Advances in understanding recessive resistance to plant viruses,” Mol Plant Pathol, vol. 5, pp. 223-233, 2004.

F. Revers, O. Le Gall, T. Candresse, and A. Maule, “New Advances in Understanding the Molecular Biology of Plant/Potyvirus Interactions,” MPMI, vol. 12, pp. 367–376, 1999.

J.C. Carrington, P.E. Jensen and M.C. Schaad, “Genetic evidence for an essential role for potyvirus CI protein in cell-to-cell movement,” Plant J, vol. 14, pp. 393-400, May 1998.

V.V. Dolja, R. Haldeman-Cahill, A.E. Montgomery, K.A. Vandenbosch and J.C. Carrington, “Capsid protein determinants involved in cell-to-cell and long distance movement of tobacco etch potyvirus,” Virology, vol. 206, pp. 1007-16, Feb 1 1995.

K.D. Kasschau, S. Cronin and J.C. Carrington, “Genome amplification and long-distance movement functions associated with the central domain of tobacco etch potyvirus helper component proteinase,” Virology, vol. 228, pp. 251-62, Feb 17 1997.

S. Vashist, L. Urena, Y. Chaudhry and I. Goodfellow, “Identification of RNAprotein interaction networks involved in the norovirus life cycle,” J Virol, vol. 86, pp. 11977-90, Nov 2012.

Z. Li and P.D. Nagy, “Diverse roles of host RNA binding proteins in RNA virus replication,” RNA Biol, vol. 8, pp. 305-15, Mar-Apr 2011.

A.L. Gutierrez-Escolano, Z.U. Brito, R.M. del Angel and X. Jiang, “Interaction of cellular proteins with the 5’ end of Norwalk virus genomic RNA,” J Virol, vol. 74, pp. 8558-62, Sep 2000.

M. Caputi and A.M. Zahler, “Determination of the RNA binding specificity of the heterogeneous nuclear ribonucleoprotein (hnRNP) H/H’/F/2H9 family,” J Biol Chem, vol. 276, pp. 43850-9, Nov 23 2001.

G.P. Patel, S. Ma and J. Bag, “The autoregulatory translational control element of poly(A)-binding protein mRNA forms a heteromeric ribonucleoprotein complex,” Nucleic Acids Res, vol. 33, pp. 7074-89, 2005.

V. Fontanes, S. Raychaudhuri and A. Dasgupta, “A cell-permeable peptide inhibits hepatitis C virus replication by sequestering IRES transacting factors,” Virology, vol. 394, pp. 82-90, Nov 10 2009.

M. Zhu, Y. Chen, X.S. Ding, S.L. Webb, T. Zhou, R.S. Nelson, et al, “Maize Elongin C interacts with the viral genome-linked protein, VPg, of Sugarcane mosaic virus and facilitates virus infection,” New Phytol, vol. 203, pp. 1291-304, Sep 2014.

Y.Q. Cheng, Z.M. Liu, J. Xu, T. Zhou, M. Wang, Y.T. Chen, et al, “HC-Pro protein of sugar cane mosaic virus interacts specifically with maize ferredoxin-5 in vitro and in planta,” J Gen Virol, vol. 89, pp. 2046-54, Aug 2008.

J. Zhang, R. Roberts and A.M. Rakotondrafara, “The role of the 5’ untranslated regions of Potyviridae in translation,” Virus Res, vol. 206, pp. 74-81, Aug 3 2015.

M.J. Adams, J.F. Antoniw and F. Beaudoin, “Overview and analysis of the polyprotein cleavage sites in the family Potyviridae,” Mol Plant Pathol, vol. 6, pp. 471-87, Jul 1 2005.

T.W. Dreher and W.A. Miller, “Translational control in positive strand RNA plant viruses,” Virology, vol. 344, pp. 185-97, Jan 5 2006.

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