Screening of a Protein That Interacts with the Matrix Attachment Region-Binding Protein fromDunaliella salina

Rui Yang,Zhaoxi Li,Yan Lin,Baosheng Yang,Tianyun Wang

doi:10.1155/2013/862450

Abstract

We isolated the matrix attachment region-binding protein (MBP) DMBP-1 from Dunaliella salina in our previous studies. MBPs are part of the cis-acting protein family cluster. The regulatory function possibly works through the interaction of the MBPs with each other. In the present study, DMBP-1 was used as the bait in screening the D. salina cDNA library for DMBP-1 interactors that could potentially mediate the DMBP-1-regulated functions. A novel MBP, namely, DMBP-2, was identified as a DMBP-1 binding partner. The cDNA of DMBP-1 was 823 bp long and contained a 573 bp open reading frame, which encoded a polypeptide of 191 amino acids. The interaction between DMBP-2 and DMBP-1 was further confirmed through glutathione S-transferase pull-down assays.

Highlights

Genome project results show that only 5% to 10% of genome DNA sequences are coding regions in the genomic DNA of all eukaryotes
Matrix attachment region DNAs (MAR) are DNA sequences that bind to nuclear matrices after being digested by restriction enzymes
We found that the MARs of Dunaliella salina can increase the transgene expression level by 1.5-fold to 4.5-fold

Summary

Introduction

Genome project results show that only 5% to 10% of genome DNA sequences are coding regions in the genomic DNA of all eukaryotes. Studying the functions of noncoding DNAs and their regulatory mechanisms is necessary. Expression regulation of the gene is performed through the interaction between cis-acting elements and transacting factors. The transacting factor must be isolated and examined to study the regulatory mechanism of a certain cis-acting element. MARs have an important role in chromatin folding and influence the adjacent endogenous gene expression as new cis-acting elements in the eukaryotic organism. MARs can function as eukaryotic organism DNA replication origins [12]. We found that the MARs of Dunaliella salina can increase the transgene expression level by 1.5-fold to 4.5-fold. We found that the regulatory function of a MAR on a transgene confers a “position effect” [13,14,15]

Methods

Results

Conclusion