Molecular biology of Lea genes of higher plants
This report contains our progress to date in determining the function of the D-7 Lea proteins in cotton embryos. We have completely sequenced the D-7 gene and established {ital E. coli} transformants which synthesize reasonable amounts of the D-7 protein. Two-dimensional electrophoresis was required to assay fractions for D-7 protein during purification to homogeneity, since D-7 has no known enzymatic activity, contains no Trp, and little Phe or Tyr, and {ital E. coli} has several proteins of similar molecular weight to D-7. Purified D-7 was used to generate monospecific antibodies which are being used for determination of the cellular distribution of D-7, and also for exact quantitation of D-7 in late-stage cotton embryos. Computerized modelling of D-7 has shown similarities to proteins with a coiled-coil structure, but fitting D-7 to this structure resulted in a violation of the handedness rule. If the pitch of the helix is changed from 3.6 to 3.667, however, a three dimensional structure (not a coiled coil) is generated which has overall energetics of formation nearly as favorable as the traditional {alpha} helix. The driving force for the change in pitch is proposed to result from favorable energetics of dimerization. Preliminary evidence indicates that D-7 does indeed dimerize in solution. Future experiments will determine the exact 3D structure of D-7 and the related protein D-29, as well as test the hypothesis that D-7 and D-29 are involved in mitigating dehydration of embryos and plants through sequestering phosphate or other ions in sufficient quantity to prevent ion precipitation or crystallization. 13 refs., 3 figs. (MHB)
- Research Organization:
- Georgia Univ., Athens, GA (United States). Research Foundation
- Sponsoring Organization:
- USDOE; USDOE, Washington, DC (United States)
- DOE Contract Number:
- FG09-89ER14071
- OSTI ID:
- 5141985
- Report Number(s):
- DOE/ER/14071-2; ON: DE92001813
- Country of Publication:
- United States
- Language:
- English
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