Abstract

In response to cold, Escherichia coli produces cold shock proteins (CSPs) that have essential roles in cold adaptation as RNA chaperones. Here, we demonstrate that Arabidopsis cold shock domain protein 3 (AtCSP3), which shares a cold shock domain with bacterial CSPs, is involved in the acquisition of freezing tolerance in plants. AtCSP3 complemented a cold-sensitive phenotype of the E. coli CSP quadruple mutant and displayed nucleic acid duplex melting activity, suggesting that AtCSP3 also functions as an RNA chaperone. Promoter-GUS transgenic plants revealed tissue-specific expression of AtCSP3 in shoot and root apical regions. When exposed to low temperature, GUS activity was extensively induced in a broader region of the roots. In transgenic plants expressing an AtCSP3-GFP fusion, GFP signals were detected in both the nucleus and cytoplasm. An AtCSP3 knock-out mutant (atcsp3-2) was sensitive to freezing compared with wild-type plants under non-acclimated and cold-acclimated conditions, whereas expression of C-repeat-binding factors and their downstream genes during cold acclimation was not altered in the atcsp3-2 mutant. Overexpression of AtCSP3 in transgenic plants conferred enhanced freezing tolerance over wild-type plants. Together, the data demonstrated an essential role of RNA chaperones for cold adaptation in higher plants.

Highlights

  • Binding factors (CBF) or dehydration responsive elementbinding protein 1 (DREB1) have been identified as transcription activators for COR gene expression [3, 4]

  • Biochemical Function of AtCSP3 Protein—AtCSP3 is comprised of a cold shock domain (CSD) with two consensus RNAbinding motifs (RNP1 and RNP2) and a glycine-rich region interspersed by seven CCHC-type zinc finger motifs (Fig. 1A)

  • BX04 transformed with the vector alone showed no visible growth at 17 °C, whereas this growth defect was suppressed by expression of AtCSP3 in BX04 (Fig. 1B)

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Summary

RNA Chaperone Regulates Freezing Tolerance

Conserved CSD is found in a diverse genera of lower and higher plants [23]. Arabidopsis thaliana has four CSD proteins that displayed differential regulation in response to low temperature [23]. Two of these proteins (AtGRP2/AtCSP2/At4g38680 and AtGRP2b/AtCSP4/At2g21060) contain two CCHC zinc fingers and the other two (AtCSP1/At4g36020 and AtCSP3/ At2g17870) contain seven CCHC zinc fingers within the glycine-rich region [23]. AtCSP2 has been subject to further characterization [27,28,29] and shown to unwind a nucleic acid duplex and partially complement the E. coli cspA, cspB, cspE, cspG quadruple deletion mutant [27]. In vivo functional analyses with overexpressors and a knock-out mutant as well as expression analyses indicate that AtCSP3 regulates freezing tolerance in Arabidopsis during cold acclimation independent of the CBF/DREB1 pathway

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