Abstract
X-arrestin (arrestin-3) is an arrestin present specifically in the outer segments of red-, green-, and blue-cone photoreceptors. The X-arrestin gene is on Xcen-q22, and consists of 17 exons with a promoter containing a TATA box and elements important for photoreceptor expression, including three CRX and one PCE-1-like element. In order to delineate the promoter structure necessary for the pan-cone-specific expression of X-arrestin, the expression of the gene in retinoblastoma cell lines was investigated, and a structure-function analysis of the promoter was conducted in the appropriate cellular substrate. Expression of X-arrestin was detected at a low level in the Y79 retinoblastoma cell line but not in the WERI retinoblastoma cell line. Truncation and expression analysis of the X-arrestin promoter in Y79 showed maximal activity in the proximal 378-bp region containing the CRX and PCE-1-like elements upstream of the TATA and CAAT boxes and a negative regulator in the distal 1-2-kbp region. Mutagenesis of the three CRX and PCE-1-like elements and expression analysis demonstrated complete elimination of the promoter activity. Mutagenesis of the TATA box and PCE-1-like element individually resulted in similar decrease in promoter activity, but the decrease in the promoter activity was greater when the CRX elements were mutagenized with a 5′ to 3′ spatial gradient in the negative effect, suggesting a cooperative effect of the three CRX elements. The regulation of expression from this promoter may involve the binding of a multi-protein enhanceosome complex at the CRX triplet and the PCE-1-like element, resulting in the recruitment and activation of the RNA polymerase II complex at the downstream TATA box.
| Original language | English |
|---|---|
| Pages (from-to) | 139-147 |
| Number of pages | 9 |
| Journal | Gene |
| Volume | 339 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Sep 15 2004 |
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Keywords
- Cone photoreceptors
- CRX, PCE-1
- Y79 retinoblastoma
ASJC Scopus subject areas
- Genetics
Cite this
Truncation and mutagenesis analysis of the human X-arrestin gene promoter. / Fujimaki, Takuro; Huang, Zhen Yong; Kitagawa, Hitoshi; Sakuma, Hitoshi; Murakami, Akira; Kanai, Atsushi; McLaren, Margaret J.; Inana, George.
In: Gene, Vol. 339, No. 1-2, 15.09.2004, p. 139-147.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Truncation and mutagenesis analysis of the human X-arrestin gene promoter
AU - Fujimaki, Takuro
AU - Huang, Zhen Yong
AU - Kitagawa, Hitoshi
AU - Sakuma, Hitoshi
AU - Murakami, Akira
AU - Kanai, Atsushi
AU - McLaren, Margaret J.
AU - Inana, George
PY - 2004/9/15
Y1 - 2004/9/15
N2 - X-arrestin (arrestin-3) is an arrestin present specifically in the outer segments of red-, green-, and blue-cone photoreceptors. The X-arrestin gene is on Xcen-q22, and consists of 17 exons with a promoter containing a TATA box and elements important for photoreceptor expression, including three CRX and one PCE-1-like element. In order to delineate the promoter structure necessary for the pan-cone-specific expression of X-arrestin, the expression of the gene in retinoblastoma cell lines was investigated, and a structure-function analysis of the promoter was conducted in the appropriate cellular substrate. Expression of X-arrestin was detected at a low level in the Y79 retinoblastoma cell line but not in the WERI retinoblastoma cell line. Truncation and expression analysis of the X-arrestin promoter in Y79 showed maximal activity in the proximal 378-bp region containing the CRX and PCE-1-like elements upstream of the TATA and CAAT boxes and a negative regulator in the distal 1-2-kbp region. Mutagenesis of the three CRX and PCE-1-like elements and expression analysis demonstrated complete elimination of the promoter activity. Mutagenesis of the TATA box and PCE-1-like element individually resulted in similar decrease in promoter activity, but the decrease in the promoter activity was greater when the CRX elements were mutagenized with a 5′ to 3′ spatial gradient in the negative effect, suggesting a cooperative effect of the three CRX elements. The regulation of expression from this promoter may involve the binding of a multi-protein enhanceosome complex at the CRX triplet and the PCE-1-like element, resulting in the recruitment and activation of the RNA polymerase II complex at the downstream TATA box.
AB - X-arrestin (arrestin-3) is an arrestin present specifically in the outer segments of red-, green-, and blue-cone photoreceptors. The X-arrestin gene is on Xcen-q22, and consists of 17 exons with a promoter containing a TATA box and elements important for photoreceptor expression, including three CRX and one PCE-1-like element. In order to delineate the promoter structure necessary for the pan-cone-specific expression of X-arrestin, the expression of the gene in retinoblastoma cell lines was investigated, and a structure-function analysis of the promoter was conducted in the appropriate cellular substrate. Expression of X-arrestin was detected at a low level in the Y79 retinoblastoma cell line but not in the WERI retinoblastoma cell line. Truncation and expression analysis of the X-arrestin promoter in Y79 showed maximal activity in the proximal 378-bp region containing the CRX and PCE-1-like elements upstream of the TATA and CAAT boxes and a negative regulator in the distal 1-2-kbp region. Mutagenesis of the three CRX and PCE-1-like elements and expression analysis demonstrated complete elimination of the promoter activity. Mutagenesis of the TATA box and PCE-1-like element individually resulted in similar decrease in promoter activity, but the decrease in the promoter activity was greater when the CRX elements were mutagenized with a 5′ to 3′ spatial gradient in the negative effect, suggesting a cooperative effect of the three CRX elements. The regulation of expression from this promoter may involve the binding of a multi-protein enhanceosome complex at the CRX triplet and the PCE-1-like element, resulting in the recruitment and activation of the RNA polymerase II complex at the downstream TATA box.
KW - Cone photoreceptors
KW - CRX, PCE-1
KW - Y79 retinoblastoma
UR - http://www.scopus.com/inward/record.url?scp=4444227235&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=4444227235&partnerID=8YFLogxK
U2 - 10.1016/j.gene.2004.06.032
DO - 10.1016/j.gene.2004.06.032
M3 - Article
C2 - 15363854
AN - SCOPUS:4444227235
VL - 339
SP - 139
EP - 147
JO - Gene
JF - Gene
SN - 0378-1119
IS - 1-2
ER -