Mutational analyses of the putative calcium binding site and hinge of the turnip crinkle virus coat protein.
The turnip crinkle carmovirus (TCV) coat protein (CP) is folded into R (RNA-binding), S (shell), and P (protruding) domains. The S domain is an eight-stranded beta barrel common to the coat protein subunits of most RNA viruses. A five-amino-acid hinge connects the S and P domains. In assembled particles, each pair of CP subunits is thought to bind a single calcium ion through interactions with three residues of one subunit and two residues of a neighboring subunit. These five residues comprise the putative calcium-binding site (CBS). The putative CBS and hinge are adjacent to one another. Mutations were introduced into the putative CBS or hinge in an effort to further determine the biological functions of TCV CP. One putative CBS mutant, TCV-M32, exhibited wild-type cell-to-cell movement but failed to move systemically in Nicotiana benthamiana, and particles were not detected. Another putative CBS mutant, TCV-M23, exhibited deficient cell-to-cell movement but particles accumulated in isolated protoplasts. Two other putative CBS mutants, TCV-M22 and -M33, showed wild-type cell-to-cell and systemic movement but elicited mild systemic symptoms that were somewhat delayed. All of the hinge mutants exhibited wild-type movement but some elicited non-wild-type symptoms. Point mutations in the putative CBS or hinge appear to alter virus-ion interactions, secondary structure, or particle conformation, thereby affecting interactions between the CP and plant hosts.
Lin
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Department of Plant Pathology, Kansas State University, Manhattan, Kansas, 66506-5502, USA.
Heaton
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LA
D001665:Binding Sites / Q000235:genetics; D001937:Brassica / Q000821:virology*; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002213:Capsid / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D019184:Carmovirus / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000235:genetics; D009154:Mutation; D011487:Protein Conformation
D002135:Calcium-Binding Proteins; D004279:DNA, Viral; D002118:Calcium