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Functional analysis of light-regulated promoter region of AtPolλ gene

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Abstract

Genetic and molecular analyses mainly in Arabidopsis and in some other plants have demonstrated involvement of light signaling in cell cycle regulation. In this report, we show light-mediated activation of the promoter of AtPolλ gene, a homolog of mammalian DNA polymerase λ in Arabidopsis thaliana and an important component of DNA damage repair/recombination machinery in plants. Analyses of the light-mediated promoter activity using various deletion versions of AtPolλ promoter in transformed Arabidopsis and tobacco (Nicotiana tabaccum) plants indicate that a 130-bp promoter region between −536 and −408 of AtPolλ promoter is essential for light-induced regulation of AtPolλ expression. DNA–protein interaction studies reveal that an ATCT-motif and AE-box light-responsive elements in the light-regulated promoter region confer light responsiveness of AtPolλ promoter. DNA-binding analysis has identified a 63-kDa trans-acting protein factor which showed specific binding to ATCT-motif, while another trans-acting factor of ~52 kDa was found to bind specifically to both ATCT and AE-box sequences. The 52-kDa protein has been identified as B3-domain transcription factor by MALDI-TOF/MS analysis. Overall, our results provide novel information on the role of light signaling in regulation of expression of an important component of DNA repair machinery in plants.

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Abbreviations

AtPolλ :

Arabidopsis thaliana DNA polymerase λ

BER:

Base excision repair

BSA:

Bovine serum albumin

DSB:

Double-strand break

GM:

Growth medium

MS:

Murashige and Skoog

NHEJ:

Non-homologous end joining

PMSF:

Phenylmethylsulfonyl fluoride

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Acknowledgments

This work was supported by the Department of Science and Technology (DST), Science and Engineering Research Council FAST TRACK research grant (Sanction No: SR/FT/LS-070/2007) to S.R. We thank Dr. Sudip Chattopadhyay, National Institute for Plant Genome Research, New Delhi, India for providing the pBI101.2 binary vector. We are thankful to Prof. John Doonan, John Innes Centre, Norwich, UK for providing the Agrobacterium strain GV3101.

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  1. Swarup Roy Choudhury

    Present address: Donald Danforth Plant Science Centre, 975 N. Warson Rd., St. Louis, MO, 63132, USA

Authors and Affiliations

  1. Department of Chemistry, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata, 700 009, West Bengal, India

    Sujit Roy & Kali Pada Das

  2. Division of Plant Biology, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata, 700 009, West Bengal, India

    Swarup Roy Choudhury & Sanjay Kumar Singh

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  1. Sujit Roy
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Correspondence to Sujit Roy.

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S. R. Choudhury contributed to this work.

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Roy, S., Choudhury, S.R., Singh, S.K. et al. Functional analysis of light-regulated promoter region of AtPolλ gene. Planta 235, 411–432 (2012). https://doi.org/10.1007/s00425-011-1517-6

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