BiP and zein binding domains within the delta zein protein.
Zeins are alcohol soluble seed storage proteins synthesized within the endosperm of maize and subsequently deposited into endoplasmic reticulum (ER) derived protein bodies. The genes encoding the beta and delta zeins were previously introduced into tobacco with the expectation of improving the nutritional quality of plants (Bagga et al. in Plant Physiol 107:13, 1997). Novel protein bodies are produced in the leaves of transgenic plants accumulating the beta or delta zein proteins. The mechanism of protein body formation within leaves is unknown. It is also unknown how zeins are retained in the ER since they do not contain known ER retention motifs. Retention may be due to an interaction of zeins with an ER chaperone such as binding luminal protein (BiP). We have demonstrated protein-protein interactions with the delta zeins, beta zeins, and BiP proteins using an E. coli two-hybrid system. In this study, four putative BiP binding motifs were identified within the delta zein protein using a BiP scoring program (Blond-Elguindi et al. in Cell 75:717, 1993). These putative binding motifs were mutated and their effects on protein interactions were analyzed in both a prokaryotic two-hybrid system and in plants. These mutations resulted in reduced BiP-zein protein interaction and also altered zein-zein interactions. Our results indicate that specific motifs are necessary for BiP-delta zein protein interactions and that there are specific motifs which are necessary for zein-zein interactions. Furthermore, our data demonstrates that zein proteins must be able to interact with BiP and zeins for their stability and ability to form protein bodies.
Randall
Jennifer J
JJ
Department of Entomology, Plant Pathology, and Weed Science, New Mexico State University, 3BE Skeen Hall, Las Cruces, NM 88003, USA. jrandall@nmsu.edu
Sutton
Dennis W
DW
Hanson
Stephen F
SF
Kemp
John D
JD
D016960:Agrobacterium tumefaciens; D020816:Amino Acid Motifs; D029681:Arabidopsis Proteins; D002352:Carrier Proteins / Q000378:metabolism*; D004926:Escherichia coli; D009154:Mutation; D018515:Plant Leaves; D010940:Plant Proteins / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011485:Protein Binding; D020798:Two-Hybrid System Techniques; D003313:Zea mays; D015028:Zein / Q000737:chemistry / Q000378:metabolism*
D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D010940:Plant Proteins; C077221:BIP protein, Arabidopsis; D015028:Zein
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