Amino Acids

, Volume 36, Issue 1, pp 137–146 | Cite as

Characterization of a histidine- and alanine-rich protein showing interaction with calreticulin in rice

Original Article
First Online:
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Abstract

Calreticulin (CRT) is a major calcium-sequestering protein in the endoplasmic reticulum and has been implicated in a variety of cellular functions. To analyze the function of CRT in rice, a yeast two-hybrid protein interaction assay was used for identifying interacting proteins. Fourteen of 17 interacting cDNA clones found coded for a novel histidine- and alanine-rich protein (OsHARP) of 342 amino acid residues. The mRNA expression level of OsHARP was up-regulated in rice seedlings treated with gibberellin (GA), but not ABA and showed a similar pattern as OsCRT mRNA. Rice plants transformed with the OsHARP promoter-GUS construct showed GUS staining in the basal parts of leaf sheaths, and although GUS activity increased when treated with GA3, it was not as high an increase as when mRNA was analyzed. To elucidate the role of OsHARP in leaf sheath elongation, antisense OsHARP transgenic rice lines were constructed. The antisense OsHARP transgenic rice plants were consistently shorter than the vector control under normal conditions. To examine whether OsHARP expression would affect other proteins, basal leaf sheaths from antisense OsHARP transgenic rice plants were analyzed using proteomic techniques. In antisense transgenic-rice OsHARP plants, OsCRT was down-regulated and the levels of 20 other proteins were changed compared to the pattern of the vector control. These results signify an important role of HARP in rice leaf sheath cell division or elongation and suggest that CRT may interact with HARP during certain stages of development.

Keywords

Calreticulin Histidine- and alanine-rich protein Leaf sheath elongation Rice 

Abbreviations

HARP

Histidine- and alanine-rich protein

CRT

Calreticulin

GA

Gibberellin

2D-PAGE

Two-dimensional polyacrylamide gel electrophoresis

CBB

Coomassie brilliant blue

PBS

Phosphate-buffered saline

PVDF

Polyvinylidene difluoride

IEF

Isoelectric focusing

MS

Mass spectrometer

BL

Brassinolide

BA

6-Benzyladenine

2,4-D

2,4-Dichlorophenoxyacetic acid

ABA

Abscisic acid

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Notes

Acknowledgments

Authors are grateful to Dr. K. Nakamura of Nagoya University for providing pIG121-Hm vector for rice transformation, and Dr. E.E.Hood of ProdiGene for providing Agrobacterium strain EHA105.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.National Institute of Crop ScienceTsukubaJapan
  2. 2.National Institute of Agrobiological SciencesTsukubaJapan

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