Lakshman D K, Jian J, Tavantzis S M
Department of Biological Sciences, University of Maine, Orono, ME 04469-5722, USA.
Proc Natl Acad Sci U S A. 1998 May 26;95(11):6425-9. doi: 10.1073/pnas.95.11.6425.
M2 is a double-stranded RNA (dsRNA) element occurring in the hypovirulent isolate Rhs 1A1 of the plant pathogenic basidiomycete Rhizoctonia solani. Rhs 1A1 originated as a sector of the virulent field isolate Rhs 1AP, which contains no detectable amount of the M2 dsRNA. The complete sequence (3,570 bp) of the M2 dsRNA has been determined. A 6.9-kbp segment of total DNA from either Rhs 1A1 or Rhs 1AP hybridizes with an M2-specific cDNA probe. The sequences of M2 dsRNA and of PCR products generated from Rhs 1A1 total DNA were found to be identical. Thus this report describes a fungal host containing full-length DNA copies of a dsRNA element. A major portion of the M2 dsRNA is located in the cytoplasm, whereas a smaller amount is found in mitochondria. Based on either the universal or the mitochondrial genetic code of filamentous fungi, one strand of M2 encodes a putative protein of 754 amino acids. The resulting polypeptide has all four motifs of a dsRNA viral RNA-dependent RNA polymerase (RDRP) and is phylogenetically related to the RDRP of a mitochondrial dsRNA associated with hypovirulence in strain NB631 of Cryphonectria parasitica, incitant of chestnut blight. This polypeptide also has significant sequence similarity with two domains of a pentafunctional polypeptide, which catalyzes the five central steps of the shikimate pathway in yeast and filamentous fungi.
M2是一种双链RNA(dsRNA)元件,存在于植物病原担子菌立枯丝核菌的低毒分离株Rhs 1A1中。Rhs 1A1起源于致病力强的田间分离株Rhs 1AP的一个扇形区域,该区域未检测到M2双链RNA。已确定M2双链RNA的完整序列(3570 bp)。来自Rhs 1A1或Rhs 1AP的总DNA的一个6.9 kbp片段与M2特异性cDNA探针杂交。发现M2双链RNA的序列与从Rhs 1A1总DNA产生的PCR产物的序列相同。因此,本报告描述了一种含有dsRNA元件全长DNA拷贝的真菌宿主。M2双链RNA的大部分位于细胞质中,而线粒体中的含量较少。根据丝状真菌的通用遗传密码或线粒体遗传密码,M2的一条链编码一种推定的754个氨基酸的蛋白质。所得多肽具有dsRNA病毒RNA依赖性RNA聚合酶(RDRP)的所有四个基序,并且在系统发育上与寄生隐孢菌NB631菌株中与低毒力相关的线粒体dsRNA的RDRP相关,NB631菌株是栗疫病的病原体。该多肽还与一种五功能多肽的两个结构域具有显著的序列相似性,该五功能多肽催化酵母和丝状真菌中莽草酸途径的五个中心步骤。