Desiccation and zinc binding induce transition of tomato abscisic acid stress ripening 1, a water stress- and salt stress-regulated plant-specific protein, from unfolded to folded state.
Abscisic acid stress ripening 1 (ASR1) is a low molecular weight plant-specific protein encoded by an abiotic stress-regulated gene. Overexpression of ASR1 in transgenic plants increases their salt tolerance. The ASR1 protein possesses a zinc-dependent DNA-binding activity. The DNA-binding site was mapped to the central part of the polypeptide using truncated forms of the protein. Two additional zinc-binding sites were shown to be localized at the amino terminus of the polypeptide. ASR1 protein is presumed to be an intrinsically unstructured protein using a number of prediction algorithms. The degree of order of ASR1 was determined experimentally using nontagged recombinant protein expressed in Escherichia coli and purified to homogeneity. Purified ASR1 was shown to be unfolded using dynamic light scattering, gel filtration, microcalorimetry, circular dichroism, and Fourier transform infrared spectrometry. The protein was shown to be monomeric by analytical ultracentrifugation. Addition of zinc ions resulted in a global change in ASR1 structure from monomer to homodimer. Upon binding of zinc ions, the protein becomes ordered as shown by Fourier transform infrared spectrometry and microcalorimetry, concomitant with dimerization. Tomato (Solanum lycopersicum) leaf soluble ASR1 is unstructured in the absence of added zinc and gains structure upon binding of the metal ion. The effect of zinc binding on ASR1 folding and dimerization is discussed.
Goldgur
Yehuda
Y
Department of Chemistry, Ben-Gurion University, Beer-Sheva 84105, Israel.
Rom
Slava
S
Ghirlando
Rodolfo
R
Shkolnik
Doron
D
Shadrin
Natalia
N
Konrad
Zvia
Z
Bar-Zvi
Dudy
D
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D003600:Cytosol / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D018551:Solanum lycopersicum / Q000187:drug effects / Q000378:metabolism*; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism*; D011058:Pollen / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D012639:Seeds / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology*; D014867:Water / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism*
C091853:Asr1 protein, Lycopersicon esculentum; D010940:Plant Proteins; D014867:Water; D012965:Sodium Chloride; D015032:Zinc