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Mol. Cells 2013; 36(2): 127-137

Published online August 31, 2013

https://doi.org/10.1007/s10059-013-0092-z

© The Korean Society for Molecular and Cellular Biology

TM6, a Novel Nuclear Matrix Attachment Region, Enhances Its Flanking Gene Expression through Influencing Their Chromatin Structure

Lusha Ji, Rui Xu, Longtao Lu, Jiedao Zhang, Guodong Yang, Jinguang Huang, Changai Wu, and Chengchao Zheng

1State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, P.R. China, 2Present address: College of Life Sciences, Liaocheng University, Liaocheng, Shandong 252059, P.R. China, 3Present address: Weifang Traditional Chinese Medicine Hospital, Weifang, Shandong 261061, P.R.China, 4These authors contributed equally to this work.

Received: March 21, 2013; Revised: May 21, 2013; Accepted: May 28, 2013

Abstract

Nuclear matrix attachment regions (MARs) regulate the higher-order organization of chromatin and affect the expression of their flanking genes. In this study, a tobacco MAR, TM6, was isolated and demonstrated to remarkably increase the expression of four different promoters that drive gusA gene and adjacent nptII gene. In turn, this expression enhanced the transformation frequency of transgenic tobacco. Deletion analysis of topoisomerase IIbinding site, AT-rich element, and MAR recognition signature (MRS) showed that MRS has the highest contribution (61.7%) to the TM6 sequence-mediated transcription activation. Micrococcal nuclease (MNase) accessibility assay showed that 35S and NOS promoter regions with TM6 are more sensitive than those without TM6. The analysis also revealed that TM6 reduces promoter DNA methylation which can affect the gusA expression. In addition, two tobacco chromatin-associated proteins, NtMBP1 and NtHMGB, isolated using a yeast one-hybrid system, specifically bound to the TM6II-1 region (761 bp to 870 bp) and to the MRS element in the TM6II-2 (934 bp to 1,021 bp) region, respectively. We thus suggested that TM6 mediated its chromatin opening and chromatin accessibility of its flanking promoters with consequent enhancement of transcription.

Keywords chromatin accessibility, DNA methylation, matrix attachment regions, transcription activation, transformation

Article

Research Article

Mol. Cells 2013; 36(2): 127-137

Published online August 31, 2013 https://doi.org/10.1007/s10059-013-0092-z

Copyright © The Korean Society for Molecular and Cellular Biology.

TM6, a Novel Nuclear Matrix Attachment Region, Enhances Its Flanking Gene Expression through Influencing Their Chromatin Structure

Lusha Ji, Rui Xu, Longtao Lu, Jiedao Zhang, Guodong Yang, Jinguang Huang, Changai Wu, and Chengchao Zheng

1State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, P.R. China, 2Present address: College of Life Sciences, Liaocheng University, Liaocheng, Shandong 252059, P.R. China, 3Present address: Weifang Traditional Chinese Medicine Hospital, Weifang, Shandong 261061, P.R.China, 4These authors contributed equally to this work.

Received: March 21, 2013; Revised: May 21, 2013; Accepted: May 28, 2013

Abstract

Nuclear matrix attachment regions (MARs) regulate the higher-order organization of chromatin and affect the expression of their flanking genes. In this study, a tobacco MAR, TM6, was isolated and demonstrated to remarkably increase the expression of four different promoters that drive gusA gene and adjacent nptII gene. In turn, this expression enhanced the transformation frequency of transgenic tobacco. Deletion analysis of topoisomerase IIbinding site, AT-rich element, and MAR recognition signature (MRS) showed that MRS has the highest contribution (61.7%) to the TM6 sequence-mediated transcription activation. Micrococcal nuclease (MNase) accessibility assay showed that 35S and NOS promoter regions with TM6 are more sensitive than those without TM6. The analysis also revealed that TM6 reduces promoter DNA methylation which can affect the gusA expression. In addition, two tobacco chromatin-associated proteins, NtMBP1 and NtHMGB, isolated using a yeast one-hybrid system, specifically bound to the TM6II-1 region (761 bp to 870 bp) and to the MRS element in the TM6II-2 (934 bp to 1,021 bp) region, respectively. We thus suggested that TM6 mediated its chromatin opening and chromatin accessibility of its flanking promoters with consequent enhancement of transcription.

Keywords: chromatin accessibility, DNA methylation, matrix attachment regions, transcription activation, transformation

Mol. Cells
Sep 30, 2022 Vol.45 No.9, pp. 603~672
COVER PICTURE
The Target of Rapamycin Complex (TORC) is a central regulatory hub in eukaryotes, which is well conserved in diverse plant species, including tomato (Solanum lycopersicum). Inhibition of TORC genes (SlTOR, SlLST8, and SlRAPTOR) by VIGS (virus-induced gene silencing) results in early fruit ripening in tomato. The red/ orange tomatoes are early-ripened TORC-silenced fruits, while the green tomato is a control fruit. Top, left, control fruit (TRV2-myc); top, right, TRV2-SlLST8; bottom, left, TRV2-SlTOR; bottom, right, TRV2-SlRAPTOR(Choi et al., pp. 660-672).

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