Mapping of a hypovirus p29 protease symptom determinant domain with sequence similarity to potyvirus HC-Pro protease

被引:51
作者
Suzuki, N [1 ]
Chen, BS [1 ]
Nuss, DL [1 ]
机构
[1] Univ Maryland, Inst Biotechnol, Agr Biotechnol Ctr, College Pk, MD 20742 USA
关键词
D O I
10.1128/JVI.73.11.9478-9484.1999
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Hypovirus infection of the chestnut blight fungus Cryphonectria parasitica results in a spectrum of phenotypic changes that can include alterations in colony morphology and significant reductions in pigmentation, asexual sporulation, and virulence (hypovirulence), Deletion of 88% [Phe(25) to Pro(243)] of the virus-encoded papain-like protease, p29, in the context of an infectious cDNA clone of the prototypic hypovirus CHV1-EP713 (recombinant virus Delta p29) partially relieved virus-mediated suppression of pigmentation and sporulation without altering the level of hypovirulence, We now report mapping of the p29 symptom determinant domain to a region extending from Phe(25) through Gln(73) by a gain-of-function analysis following progressive repair of the Delta p29 deletion mutant. This domain was previously shown to share sequence similarity [including conserved cysteine residues Cys(38), Cys(48), Cys(70), and Cys(72)] with the N-terminal portion of the potyvirus-encoded helper component-proteinase (HC-Pro), a multifunctional protein implicated in aphid-mediated transmission, genome amplification, polyprotein processing, long-distance movement, and suppression of posttranscriptional silencing, Substitution of a glycine residue for either Cys(38) or Cys(48) resulted in no qualitative or quantitative changes in virus-mediated symptoms. Unexpectedly, mutation of Cys(70) resulted in a very severe phenotype that included significantly reduced mycelial growth and profoundly altered colony morphology. In contrast, substitution for Cys(72) resulted in a less severe symptom phenotype approaching that observed for Delta p29, The finding that p29-mediated symptom expression is influenced by two cysteine residues that are conserved in the potyvirus-encoded HC-Pro raises the possibility that these related viral-papain-like proteases function in their respective fungal and plant hosts by impacting ancestrally related regulatory pathways.
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页码:9478 / 9484
页数:7
相关论文
共 31 条
[1]   BIOLOGICAL-CONTROL OF CHESTNUT BLIGHT [J].
ANAGNOSTAKIS, SL .
SCIENCE, 1982, 215 (4532) :466-471
[2]   MUTATIONAL ANALYSIS OF THE HELPER COMPONENT-PROTEINASE GENE OF A POTYVIRUS - EFFECTS OF AMINO-ACID SUBSTITUTIONS, DELETIONS, AND GENE REPLACEMENT ON VIRULENCE AND APHID TRANSMISSIBILITY [J].
ATREYA, CD ;
PIRONE, TP .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (24) :11919-11923
[3]   SITE-DIRECTED MUTATIONS IN THE POTYVIRUS HC-PRO GENE AFFECT HELPER COMPONENT ACTIVITY, VIRUS ACCUMULATION, AND SYMPTOM EXPRESSION IN INFECTED TOBACCO PLANTS [J].
ATREYA, CD ;
ATREYA, PL ;
THORNBURY, DW ;
PIRONE, TP .
VIROLOGY, 1992, 191 (01) :106-111
[4]   A 2ND PROTEINASE ENCODED BY A PLANT POTYVIRUS GENOME [J].
CARRINGTON, JC ;
CARY, SM ;
PARKS, TD ;
DOUGHERTY, WG .
EMBO JOURNAL, 1989, 8 (02) :365-370
[5]   ATTENUATION OF FUNGAL VIRULENCE BY SYNTHETIC INFECTIOUS HYPOVIRUS TRANSCRIPTS [J].
CHEN, BS ;
CHOI, GH ;
NUSS, DL .
SCIENCE, 1994, 264 (5166) :1762-1764
[6]   Infectious cDNA clone of hypovirus CHV1-Euro7:: a comparative virology approach to investigate virus-mediated hypovirulence of the chestnut blight fungus Cryphonectria parasitica [J].
Chen, BS ;
Nuss, DL .
JOURNAL OF VIROLOGY, 1999, 73 (02) :985-992
[7]   Phenotypic changes associated with wild-type and mutant hypovirus RNA transfection of plant pathogenic fungi phylogenetically related to Cryphonectria parasitica [J].
Chen, BS ;
Chen, CH ;
Bowman, BH ;
Nuss, DL .
PHYTOPATHOLOGY, 1996, 86 (03) :301-310
[8]   HYPOVIRULENCE OF CHESTNUT BLIGHT FUNGUS CONFERRED BY AN INFECTIOUS VIRAL CDNA [J].
CHOI, GH ;
NUSS, DL .
SCIENCE, 1992, 257 (5071) :800-803
[9]   A VIRAL GENE CONFERS HYPOVIRULENCE-ASSOCIATED TRAITS TO THE CHESTNUT BLIGHT FUNGUS [J].
CHOI, GH ;
NUSS, DL .
EMBO JOURNAL, 1992, 11 (02) :473-477
[10]   THE AUTOCATALYTIC PROTEASE-P-29 ENCODED BY A HYPOVIRULENCE-ASSOCIATED VIRUS OF THE CHESTNUT BLIGHT FUNGUS RESEMBLES THE POTYVIRUS-ENCODED PROTEASE HC-PRO [J].
CHOI, GH ;
PAWLYK, DM ;
NUSS, DL .
VIROLOGY, 1991, 183 (02) :747-752