Prokaryotic Expression of Rice Ospgip1 Gene and Bioinformatic Analysis of Encoded Product

Abstract

Abstract: Using the reference sequences of pgip genes in GenBank, a fragment of 930 bp covering the open reading frame (ORF) of rice Ospgip1 (Oryza sativa polygalacturonase-inhibiting protein 1) was amplified. The prokaryotic expression product of the gene inhibited the growth of Rhizoctonia solani, the causal agent of rice sheath blight, and reduced its polygalacturonase activity. Bioinformatic analysis showed that OsPGIP1 is a hydrophobic protein with a molecular weight of 32.8 kDa and an isoelectric point (pI) of 7.26. The protein is mainly located in the cell wall of rice, and its signal peptide cleavage site is located between the 17th and 18th amino acids. There are four cysteines in both the N- and C-termini of the deduced protein, which can form three disulfide bonds (between the 56th and 63rd, the 278th and 298th, and the 300th and 308th amino acids). The protein has a typical leucine-rich repeat (LRR) domain, and its secondary structure comprises α-helices, β-sheets and irregular coils. Compared with polygalacturonase-inhibiting proteins (PGIPs) from other plants, the 7th LRR is absent in OsPGIP1. The nine LRRs could form a cleft that might associate with proteins from pathogenic fungi, such as polygalacturonase.

Key words: Ospgip1 gene, polygalacturonase-inhibiting protein, prokaryotic expression, bioinformatic analysis, rice, Rhizoctonia solani

CHEN Xi-jun, LIU Xiao-wei, ZUO Si-min, MA Yu-yin, TONG Yun-hui1 PAN Xue-biao, XU Jing-you. Prokaryotic Expression of Rice Ospgip1 Gene and Bioinformatic Analysis of Encoded Product[J]. RICE SCIENCE, 2011, 18(4): 250-256.

References

Bendtsen J D, Nielsen H, von Heijne G V, Brunak S. 2004. Improved prediction of signal peptides:SignalP 3.0. J Mol Biol, 340(4): 783?795.
Blom N, Gammeltoft S, Brunak S. 1999. Sequence and structure-based prediction of eukaryotic protein phosphorylation sites. J Mol Biol, 294(5): 1351?1362.
Chen X J, Wang Y D, Zuo S M, Tong Y H, Pan X B, Xu J Y. 2010. Isolation, purification and characterization of polygalacturonase (PG) from Rhizoctonia solani, the pathogen of rice sheath blight. Acta Phytopathologica sinica, 40(3): 276?281. (in Chinese with English abstract)
Chen X J, Zhang H, Tong Y H, Xu J Y. 2006. Cell wall degrading enzymes produced by Rhizoctonia solani and their pathogenicity to rice plants. Jiangsu J Agric Sci, 22(1): 24?28. (in Chinese with English abstract)
Cheng J, Randall A Z, Sweredoski M J, Baldi P. 2005. SCRATCH: A protein structure and structural feature prediction server. Nucl Acids Res, 33(7): 72?76.
Clark E D B. 1998. Refolding of recombinant proteins. Curr Opin Biotechnol, 9: 157?163.
Cserz? M, Eisenhaber F, Eisenhaber B, Simon I. 2002. On filtering false positive transmembrane protein predictions. Protein Eng, 15(9): 745?752.
D’Ovidio R, Roberti S, Di Giovanni M, Capodicasa C, Melaragni M, Sella L, Tosi P, Favaron F. 2006. The characterization of the soybean polygalacturonase-inhibiting proteins (Pgip) gene family reveals that a single member is responsible for the activity detected in soybean tissues. Planta, 224(3): 633?645.
De Lorenzo G, D’Ovidio R, Cervone F. 2001. The role of polygacturonase-inhibiting proteins (PGIPs) in defense against pathogenic fungi. Annu Rev Phytopathol, 39: 313?335.
Di Matteo A, Federici L, Mattei B, Salvi G, Johnson K A. 2003. The crystal structure of PGIP (polygalacturonase-inhibiting protein), a leucine-rich repeat protein involved in plant defense. Proc Natl Acad Sci USA, 100: 10124?10128.
Favaron F. 2001. Gel detection of Allium porrum polygalacturonase-inhibiting protein reveals a high number of isoforms. Physiol Mol Plant Pathol, 58: 239?245.
Federici L, Di Matteo A, Fernandez-Recio J, Tsernoglou D, Cervone F. 2006. Polygalacturonases inhibiting proteins: Players in plant innate immunity? Trends Plant Sci, 11(2): 65?70.
Fong J H, Keating A E, Singh M. 2004. Predicting specificity in bZIP coiled-coil protein interactions. Genome Biol, 5(2): 2?10.
Harrison R G. 2000. Expression of soluble heterologous proteins via fusion with NusA protein. inNovation, 11: 4?7.
James J T, Dubery I A. 2001. Inhibition of polygalacturonase from Verticillium dahliae by a polygalacturonase inhibiting protein from cotton. Phytochemistry, 57: 149?156.
Janni M, Di Giovanni M, Roberti S, Capodicasa C, D’Ovidio R. 2006. Characterization of expressed pgip genes in rice and wheat reveals similar extent of sequence variation to dicot PGIPs and identifies an active PGIP lacking an entire LRR repeat. Theor Appl Genet, 113: 1233?1245.
Janni M, Sella L, Favaron F, Blechl A E, De Lorenzo G, D'Ovidio R. 2008. The expression of a bean PGIP in transgenic wheat confers increased resistance to fungal pathogen Bipolaris sorokiniana. Am Phytopathol Soc, 21(2): 171?177.
Khanuja S P S, Shasany A K, Darokar M P, Kumar S. 1999. Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Mol Biol Rep, 17: 1?7.
Lambert C, Léonard N, De Bolle X, Depiereux E. 2002. ESyPred3D: Prediction of proteins 3D structures. Bioinformatics, 18(9): 1250?1256.
Leckie F, Mattei B, Capodicasa C, Hemmings A, Nuss L, Aracri B, De Lorenzo G, Cervone F. 1999. The specificity of polygalacturonase-inhibiting protein (PGIP): A single amino acid substitution in the solvent-exposed β-strand/β-turn region of the leucine-rich repeats (LRRs) confers a new recognition capability. EMBO J, 18(9): 2352?2363.
Li R, Rimmer R, Yu M, Sharpe A G, Séguin-Swartz G, Lydiate D, Hegedus D D. 2003.Two Brassica napus polygalacturonase inhibitory protein genes are expressed at different levels in response to biotic and abiotic stresses. Planta, 217(2): 299?308.
Liao H N, Xiao L S, Wang H S. 1997.Analysis of sheath blight developing history and evolving matter. Guangxi Plant Prot, (3): 35?38. (in Chinese)
Nakao M C, Nakai K. 2002. Improvement of PSORT II protein sorting prediction for mammalian proteins. Genome Informatics, 13: 441?442.
Oeser B, Heidrich P M, Müller U, Tudzynski P, Tenberge K B. 2002. Polygalacturonase is a pathogenicity factor in the Claviceps purpurea/rye interaction. Fung Genet Biol, 36(3): 176?186.
Roco A, Casta?eda P, Pérez L M. 1993. Oligosaccharides released by pectinase treatment of Citrus limon seedlings are elicitors of the plant response. Phytochemistry, 33(6): 1301?1306.
Sicilia F, Fernandez-Recio J, Caprari C, De Lorenzo G, Tsernoglou D, Cervone F, Federici L. 2005. The polygalacturonase- inhibiting protein PGIP2 of Phaseolus vulgaris has evolved a mixed mode of inhibition of endo-polygalacturonase PG1 of Botrytis cinerea. Plant Physiol, 139(3): 1380?1388.
Spinelli F, Mariotti L, Mattei B. Salvi G, Cervone F, Caprari C. 2009. Three aspartic acid residues of polygalacturonase-inhibiting protein (PGIP) from Phaseolus vulgaris are critical for inhibition of Fusarium phyllophilum PG. Plant Biol, 11(5): 738?743.
Stotz H U, Contos J J A, Powell A L T, Bennett A B, Labavitch J M. 1994. Structure and expression of an inhibitor of fungal polygalacturonases from tomato. Plant Mol Biol, 25(4): 607?617.
Walker J M. 2005. The Proteomics Protocol Handbook. Totow: Humana Press: 571?607.
Walton J D. 1994. Deconstructing the cell wall. Plant Physiol, 104(4): 1113?1118.
Zhang M, Guo L Q, Guan X Y, Zhang F. 2008. Study on inclusion body’s denaturation and renaturation of AiiA fusion protein. J Fujian Normal Univ:Nat Sci, 24(4): 76?79. (in Chinese with English abstract)
Zuo S M, Zhang Y F, Chen Z X, Pan X B. 2006. Establishment and inprovement of inoculation technique and rating system in researching rice sheath blight resistance in field. J Yangzhou Univ: Agric & Life Sci, 27(4): 57?61. (in Chinese with English abstract)

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