Analysis of Regulatory Regions of Emilin1 Gene and Their Combinatorial Contribution to Tissue-specific Transcription

Abstract

The location of regions that regulate transcription of the murine Emilin1 gene was investigated in a DNA fragment of 16.8 kb, including the entire gene and about 8.7 and 0.6 kb of 5-and 3-flanking sequences, respectively. The 8.7-kb segment contains the 5-end of the putative 2310015E02Rik gene and the sequence that separates it from Emilin1, whereas the 0.6-kb fragment covers the region between Emilin1 and Ketohexokinase genes. Sequence comparison between species identified several conserved regions in the 5-flanking sequence. Most of them contained chromatin DNase I-hypersensitive sites, which were located at about ؊950 (HS1), ؊3100 (HS2), ؊4750 (HS3), and ؊5150 (HS4) in cells expressing Emilin1 mRNA. Emilin1 transcription initiates at multiple sites, the major of which correspond to two Initiator sequences. Promoter assays suggest that core promoter activity was mainly dependent on Initiator1 and on Sp1-binding sites close to the Initiators. Moreover, one important regulatory region was contained between ؊1 and ؊169 bp and a second one between ؊630 bp and ؊1.1 kb. The latter harbors a putative binding site for transcription factor AP1 matching the location of HS1. The function of different regions was studied by expressing lacZ constructs in transgenic mice. The results show that the 16.8-kb segment contains regulatory sequences driving high level transcription in all the tissues where Emilin1 is expressed. Moreover, the data suggest that transcription in different tissues is achieved through combinatorial cooperation between various regions, rather than being dependent on a single cis-activating region specific for each tissue.

Vascular Smooth Muscle Emilin-1 Is a Regulator of Arteriolar Myogenic Response and Blood Pressure

“…The expression of the transgenic Emilin1 promoter in embryos reproduced the complexity expected from the distribution of endogenous gene products ( Figure I in the online-only Data Supplement). 1,3,9 Matching of this promoter activity with that of the endogenous gene was investigated in mice harboring the human EMILIN1 cDNA in Emilin1 +/− embryos. As shown in Figure Table).…”

Emilin1 gene and essential hypertension: a two-stage association study in northern Han Chinese population

“…The gC1q domain of Emilin1 can form relatively stable and compact homotrimers, and then be followed by a multimeric assembly of disulfide-bonded protomers. The transcription of Emilin1 in different tissues is probably achieved through combinatorial cooperation between various regions, rather than being dependent on a single cis-activating region specific for each tissue [27]. Emilin1 code for a secreted proteins whose hallmark is the presence of a cysteine-rich motif defined as EMI domain [28] that followed by a coiled-coil, a short collagenous domain, and a C1q-like repeat at the C terminus which has been shown that the C-terminal domain is involved in the oligomerization of Emilin1, cell adhesion and modulating growth factors' activity [29].…”

Preparation and Characterization of a Novel Monoclonal Antibody Specific to Human EMILIN-5 Protein

“…Recently, EMI domain has been found related to mesenchymal commitment during an early stage of skeletal development, indicating that EMILIN-5 might also contribute to skeletal development. (2,3) EMILIN-5 was found expressed in human mesenchymal stem cells. (1) Thus far, however, the functions of EMILIN-5 remain largely unknown.…”