Author(s): Hyun-Il Shin | Kwang-Hee Chae | Tae-Ju Cho
Journal: BMB Reports
ISSN 1976-6696
Volume: 46;
Issue: 10;
Start page: 495;
Date: 2013;
Original page
Keywords: Coat protein | Modification | RNA packaging | TYMV | Virion assembly
ABSTRACT
Turnip yellow mosaic virus (TYMV) is a positive strand RNAvirus. We have modified TYMV coat protein (CP) by inserting ac-Myc epitope peptide at the N- or C-terminus of the CP, andhave examined its effect on assembly. We introduced therecombinant CP constructs into Nicotiana benthamiana leavesby agroinfiltration. Examination of the leaf extracts by agarosegel electrophoresis and Western blot analysis showed that theCP modified at the N-terminus produced a band co-migratingwith wild-type virions. With C-terminal modification, however,the detected bands moved faster than the wild-type virions. Tofurther examine the effect, TYMV constructs producing themodified CPs were prepared. With N-terminal modification,viral RNAs were protected from RNase A. In contrast, the viralRNAs were not protected with C-terminal modification.Overall, the results suggest that virion assembly and RNApackaging occur properly when the N-terminus of CP ismodified, but not when the C-terminus is modified. [BMBReports 2013; 46(10): 495-500]
Journal: BMB Reports
ISSN 1976-6696
Volume: 46;
Issue: 10;
Start page: 495;
Date: 2013;
Original page
Keywords: Coat protein | Modification | RNA packaging | TYMV | Virion assembly
ABSTRACT
Turnip yellow mosaic virus (TYMV) is a positive strand RNAvirus. We have modified TYMV coat protein (CP) by inserting ac-Myc epitope peptide at the N- or C-terminus of the CP, andhave examined its effect on assembly. We introduced therecombinant CP constructs into Nicotiana benthamiana leavesby agroinfiltration. Examination of the leaf extracts by agarosegel electrophoresis and Western blot analysis showed that theCP modified at the N-terminus produced a band co-migratingwith wild-type virions. With C-terminal modification, however,the detected bands moved faster than the wild-type virions. Tofurther examine the effect, TYMV constructs producing themodified CPs were prepared. With N-terminal modification,viral RNAs were protected from RNase A. In contrast, the viralRNAs were not protected with C-terminal modification.Overall, the results suggest that virion assembly and RNApackaging occur properly when the N-terminus of CP ismodified, but not when the C-terminus is modified. [BMBReports 2013; 46(10): 495-500]
