A novel plant homeodomain protein interacts in a functionally relevant manner with a virus movement protein.
Tomato bushy stunt virus and its cell-to-cell movement protein (MP; P22) provide valuable tools to study trafficking of macromolecules through plants. This study shows that wild-type P22 and selected movement-defective P22 amino acid substitution mutants were equivalent for biochemical features commonly associated with MPs (i.e. RNA binding, phosphorylation, and membrane partitioning). This generated the hypothesis that their movement defect was caused by improper interaction between the P22 mutants and one or more host factors. To test this, P22 was used as bait in a yeast (Saccharomyces cerevisiae) two-hybrid screen with a tobacco (Nicotiana tabacum) cDNA library, which identified a new plant homeodomain leucine-zipper protein that reproducibly interacted with P22 but not with various control proteins. These results were confirmed with an independent in vitro binding test. An mRNA for the host protein was detected in plants, and its accumulation was enhanced upon Tomato bushy stunt virus infection of two plant species. The significance of this interaction was further demonstrated by the failure of the homeodomain protein to interact efficiently with two of the well-defined movement-deficient P22 mutants in yeast and in vitro. This is the first report, to our knowledge, that a new plant homeodomain leucine-zipper protein interacts specifically and in a functionally relevant manner with a plant virus MP.
Department of Plant Pathology and Microbiology, Texas A&M University, 2132 TAMU, College Station, Texas 77843, USA.
Bénédicte
B
Desvoyes
Sandrine
S
Faure-Rabasse
Min-Huei
MH
Chen
Jong-Won
JW
Park
Herman B
HB
Scholthof
D000595:Amino Acid Sequence; D001692:Biological Transport / Q000502:physiology; D002352:Carrier Proteins / Q000502:physiology; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D016350:Leucine Zippers / Q000502:physiology; D008969:Molecular Sequence Data; D009154:Mutation; D025941:Protein Interaction Mapping; D012333:RNA, Messenger / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014026:Nicotiana / Q000235:genetics / Q000821:virology; D017864:Tombusvirus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D002352:Carrier Proteins; D018398:Homeodomain Proteins; D012333:RNA, Messenger; D014764:Viral Proteins