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Mol. Cells 2011; 32(1): 67-76

Published online July 31, 2011

https://doi.org/10.1007/s10059-011-1031-5

© The Korean Society for Molecular and Cellular Biology

A Strong Promoter Activity of Pre-B Cell Stage-Specific Crlz1 Gene Is Caused by One Distal LEF-1 and Multiple Proximal Ets Sites

Sung-Kyun Park, Youngsook Son, and Chang-Joong Kang*

Graduate School of Biotechnology, Institute of Life Science and Resources, Kyung Hee University, Yongin 446-701, Korea

Correspondence to : *Correspondence: cjkang@khu.ac.kr

Received: February 21, 2011; Revised: April 9, 2011; Accepted: April 15, 2011

Abstract

The promoter of pre-B cell stage-specific Crlz1 gene, whose protein translocates the cytoplasmic core binding factorα(CBFβ) into the nucleus and thereby allows its heterodi-merization with Runx, has a very strong activity, which is about 25% of cytomegalovirus (CMV) promoter activity and comparable to the EF-1α promoter activity. Its transcription start site was mapped at 155 nt upstream of translation initiation codon. 5'-truncation analysis of charged amino acids rich leucine zipper 1 (Crlz1) promoter revealed that one distal region from -612 to -536 and one proximal region from -198 to -100 as numbered from the transcription start site were critical for the promoter activity. The 3'-truncation analysis of the promoter revealed that the basal promoter sequence around the transcription start site, which should be necessary for the assembly of transcription initiation complex and the start of RNA polymerase II, was also essential, although not sufficient by itself. When transcription factorαbinding sites within those two critical regions were searched by in vivo footprinting, one distal LEF-1 and multiple proximal Ets consensus-like sites were found to be footprinted. Indeed, the protein causing a footprint over the distal region was found to be LEF-1, and the ones causing three footprints over the proximal region were found to be such Ets family members as Fli-1 and GABP, as verified by EMSA and ChIP analyses. Furthermore, those LEF-1 and Ets sites were shown to drive additively a strong transcrip-tion of Crlz1 gene.

Keywords CBFβ, Crlz1, Ets, LEF-1, Runx

Article

Research Article

Mol. Cells 2011; 32(1): 67-76

Published online July 31, 2011 https://doi.org/10.1007/s10059-011-1031-5

Copyright © The Korean Society for Molecular and Cellular Biology.

A Strong Promoter Activity of Pre-B Cell Stage-Specific Crlz1 Gene Is Caused by One Distal LEF-1 and Multiple Proximal Ets Sites

Sung-Kyun Park, Youngsook Son, and Chang-Joong Kang*

Graduate School of Biotechnology, Institute of Life Science and Resources, Kyung Hee University, Yongin 446-701, Korea

Correspondence to:*Correspondence: cjkang@khu.ac.kr

Received: February 21, 2011; Revised: April 9, 2011; Accepted: April 15, 2011

Abstract

The promoter of pre-B cell stage-specific Crlz1 gene, whose protein translocates the cytoplasmic core binding factorα(CBFβ) into the nucleus and thereby allows its heterodi-merization with Runx, has a very strong activity, which is about 25% of cytomegalovirus (CMV) promoter activity and comparable to the EF-1α promoter activity. Its transcription start site was mapped at 155 nt upstream of translation initiation codon. 5'-truncation analysis of charged amino acids rich leucine zipper 1 (Crlz1) promoter revealed that one distal region from -612 to -536 and one proximal region from -198 to -100 as numbered from the transcription start site were critical for the promoter activity. The 3'-truncation analysis of the promoter revealed that the basal promoter sequence around the transcription start site, which should be necessary for the assembly of transcription initiation complex and the start of RNA polymerase II, was also essential, although not sufficient by itself. When transcription factorαbinding sites within those two critical regions were searched by in vivo footprinting, one distal LEF-1 and multiple proximal Ets consensus-like sites were found to be footprinted. Indeed, the protein causing a footprint over the distal region was found to be LEF-1, and the ones causing three footprints over the proximal region were found to be such Ets family members as Fli-1 and GABP, as verified by EMSA and ChIP analyses. Furthermore, those LEF-1 and Ets sites were shown to drive additively a strong transcrip-tion of Crlz1 gene.

Keywords: CBFβ, Crlz1, Ets, LEF-1, Runx

Mol. Cells
Jan 31, 2023 Vol.46 No.1, pp. 1~67
COVER PICTURE
RNAs form diverse shapes and play multiple functions as central molecules of gene expression. In this special issue on RNA, seven minireviews illustrate how basic concepts and recent RNA biology findings are transformed into new and exciting RNA therapeutics.

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