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Plant Physiol, February 2001, Vol. 125, pp. 797-809
Homeodomain Leucine Zipper Proteins Bind to the Phosphate
Response Domain of the Soybean VspB Tripartite
Promoter1
Zhijun
Tang,
Avi
Sadka,
Daryl T.
Morishige, and
John E.
Mullet*
Genentech, Incorporated, 1 DNA Way, Mail Stop 37, South San Francisco, California 94080 (Z.T.); Institute of
Horticulture, Agricultural Research Organization, The Volcani
Center, P.O. Box 6, Bet-Dagan 50250, Israel (A.S.); and Department of
Biochemistry and Biophysics, Texas A&M University, College Station,
Texas 77840 (D.T.M., J.E.M.)
The soybean (Glycine max L. Merr. cv Williams
82) genes VspA and VspB encode
vacuolar glycoprotein acid phosphatases that serve as vegetative
storage proteins during seed fill and early stages of seedling growth.
VspB expression is activated by jasmonates (JAs) and
sugars and down-regulated by phosphate and auxin. Previous promoter
studies demonstrated that VspB promoter sequences
between  585 and 535 mediated responses to JA, and sequences between 535 and 401 mediated responses to sugars, phosphate, and auxin. In
this study, the response domains were further delineated using transient expression of VspB
promoter- -glucuronidase constructs in tobacco protoplasts.
Sequences between 536 and 484 were identified as important for
phosphate responses, whereas the region from 486 to 427 mediated
sugar responses. Gel-shift and deoxyribonuclease-I footprinting assays
revealed four DNA-binding sites between 611 and 451 of the soybean
VspB promoter: one in the JA response domain, two in the
phosphate response domain, and one binding site in the sugar response
domain. The sequence CATTAATTAG present in the phosphate response
domain binds soybean homeodomain leucine zipper proteins, suggesting a
role for these transcription factors in phosphate-modulated gene expression.
1
This work was supported by the National Science
Foundation Genetics Program (grant no. MCB-9514034 to J.E.M.) and by
the Texas Agricultural Experiment Station.
*
Corresponding author; e-mail jmullet{at}tamu.edu; fax
409-845-9274.
© 2001 American Society of Plant Physiologists
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