scholarly journals Crystal structures of MS2 coat protein mutants in complex with wild- type RNA operator fragments

1998 ◽  
Vol 26 (5) ◽  
pp. 1345-1351 ◽  
Author(s):  
S. van den Worm
1999 ◽  
Vol 89 (7) ◽  
pp. 540-545 ◽  
Author(s):  
R. G. Guevara-González ◽  
P. L. Ramos ◽  
R. F. Rivera-Bustamante

The role of the pepper huasteco virus (PHV) coat protein (CP) gene during the infection was investigated in three different hosts by using mutations that produced truncated proteins and by complementation assays in transgenic plants. The infectivity analysis revealed that mutants that express truncated CP (CP7 and CP191) behave like the wild-type virus when inoculated onto pepper and Nicotiana benthamiana plants in terms of symptom expression and viral DNA movement. On the contrary, the CP7 mutant was unable to systemically infect tobacco plants, whereas only 10% of the plants inoculated with the CP191 mutant became infected. The CP7 mutant was complemented by coinoculating it with another geminivirus (taino tomato mottle virus). No complementation was observed in plants from nine transgenic tobacco lines expressing CP under the control of the cauliflower mosaic virus (CaMV) 35S promoter. However, 3 out of 10 lines expressing CP under the control of its own promoter (693 nucleotides) were able to complement the CP7 mutant. Interestingly, upon infection, the levels of CP mRNA in 693CP plants increased dramatically, probably due to transactivation of the CP promoter by the viral protein AC2.


2000 ◽  
Vol 298 (1) ◽  
pp. 149-162 ◽  
Author(s):  
Yafei Huang ◽  
Junichi Komoto ◽  
Kiyoshi Konishi ◽  
Yoshimi Takata ◽  
Hirofumi Ogawa ◽  
...  

2001 ◽  
Vol 82 (11) ◽  
pp. 2827-2836 ◽  
Author(s):  
Chu-Hui Chiang ◽  
Ju-Jung Wang ◽  
Fuh-Jyh Jan ◽  
Shyi-Dong Yeh ◽  
Dennis Gonsalves

Transgenic papaya cultivars SunUp and Rainbow express the coat protein (CP) gene of the mild mutant of papaya ringspot virus (PRSV) HA. Both cultivars are resistant to PRSV HA and other Hawaii isolates through homology-dependent resistance via post-transcriptional gene silencing. However, Rainbow, which is hemizygous for the CP gene, is susceptible to PRSV isolates from outside Hawaii, while the CP-homozygous SunUp is resistant to most isolates but susceptible to the YK isolate from Taiwan. To investigate the role of CP sequence similarity in overcoming the resistance of Rainbow, PRSV HA recombinants with various CP segments of the YK isolate were constructed and evaluated on Rainbow, SunUp and non-transgenic papaya. Non-transgenic papaya were severely infected by all recombinants, but Rainbow plants developed a variety of symptoms. On Rainbow, a recombinant with the entire CP gene of YK caused severe symptoms, while recombinants with only partial YK CP sequences produced a range of milder symptoms. Interestingly, a recombinant with a YK segment from the 5′ region of the CP gene caused very mild, transient symptoms, whereas recombinants with YK segments from the middle and 3′ parts of the CP gene caused prominent and lasting symptoms. SunUp was resistant to all but two recombinants, which contained the entire CP gene or the central and 3′-end regions of the CP gene and the 3′ non-coding region of YK, and the resulting symptoms were mild. It is concluded that the position of the heterologous sequences in the recombinants influences their pathogenicity on Rainbow.


2018 ◽  
Vol 124 ◽  
pp. 214-220 ◽  
Author(s):  
Eszter Szabo ◽  
Reka Mizsei ◽  
Piotr Wilk ◽  
Zsofia Zambo ◽  
Beata Torocsik ◽  
...  

Biochemistry ◽  
1995 ◽  
Vol 34 (38) ◽  
pp. 12388-12397 ◽  
Author(s):  
Amir R. Khan ◽  
Karen A. Williams ◽  
Joan M. Boggs ◽  
Charles M. Deber

2008 ◽  
Vol 191 (4) ◽  
pp. 1211-1219 ◽  
Author(s):  
Lian-Hua Xu ◽  
Shinya Fushinobu ◽  
Haruo Ikeda ◽  
Takayoshi Wakagi ◽  
Hirofumi Shoun

ABSTRACT The polyene macrolide antibiotic filipin is widely used as a probe for cholesterol in biological membranes. The filipin biosynthetic pathway of Streptomyces avermitilis contains two position-specific hydroxylases, C26-specific CYP105P1 and C1′-specific CYP105D6. In this study, we describe the three X-ray crystal structures of CYP105P1: the ligand-free wild-type (WT-free), 4-phenylimidazole-bound wild-type (WT-4PI), and ligand-free H72A mutant (H72A-free) forms. The BC loop region in the WT-free structure has a unique feature; the side chain of His72 within this region is ligated to the heme iron. On the other hand, this region is highly disordered and widely open in WT-4PI and H72A-free structures, respectively. Histidine ligation of wild-type CYP105P1 was not detectable in solution, and a type II spectral change was clearly observed when 4-phenylimidazole was titrated. The H72A mutant showed spectroscopic characteristics that were almost identical to those of the wild-type protein. In the H72A-free structure, there is a large pocket that is of the same size as the filipin molecule. The highly flexible feature of the BC loop region of CYP105P1 may be required to accept a large hydrophobic substrate.


2004 ◽  
Vol 149 (4) ◽  
pp. 699-712 ◽  
Author(s):  
C. A. Malpica ◽  
M.-E. Aleman-Verdaguer ◽  
D. M. Stark ◽  
C. M. Fauquet ◽  
R. N. Beachy ◽  
...  

1999 ◽  
Vol 12 (2) ◽  
pp. 153-162 ◽  
Author(s):  
Marise Borja ◽  
Teresa Rubio ◽  
Herman B. Scholthof ◽  
Andrew O. Jackson

Nicotiana benthamiana plants transformed with the coat protein gene of tomato bushy stunt virus (TBSV) failed to elicit effective virus resistance when inoculated with wild-type virus. Subsequently, R1 and R2 progeny from 13 transgenic lines were inoculated with a TBSV mutant containing a defective coat protein gene. Mild symptoms typical of those elicited in nontransformed plants infected with the TBSV mutant initially appeared. However, within 2 to 4 weeks, up to 20% of the transgenic plants sporadically began to develop the lethal syndrome characteristic of wild-type virus infections. RNA hybridization and immunoblot analyses of these plants and nontransformed N. benthamiana inoculated with virus from the transgenic lines indicated that wild-type virus had been regenerated by a double recombination event between the defective virus and the coat protein transgene. Similar results were obtained with a TBSV deletion mutant containing a nucleotide sequence marker, and with a chimeric cucumber necrosis virus (CNV) containing the defective TBSV coat protein gene. In both cases, purified virions contained wild-type TBSV RNA or CNV chimeric RNA derived by recombination with the transgenic coat protein mRNA. These results thus demonstrate that recombinant tombusviruses can arise frequently from viral genes expressed in transgenic plants.


2002 ◽  
Vol 277 (37) ◽  
pp. 34499-34507 ◽  
Author(s):  
Andrew P. Yeh ◽  
Xavier I. Ambroggio ◽  
Susana L. A. Andrade ◽  
Oliver Einsle ◽  
Claire Chatelet ◽  
...  

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