Truncation and mutagenesis analysis of the human X-arrestin gene promoter

Takuro Fujimaki, Zhen Yong Huang, Hitoshi Kitagawa, Hitoshi Sakuma, Akira Murakami, Atsushi Kanai, Margaret J. McLaren, George Inana

Research output: Contribution to journalArticle

4 Citations (Scopus)

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 languageEnglish
Pages (from-to)139-147
Number of pages9
JournalGene
Volume339
Issue number1-2
DOIs
StatePublished - Sep 15 2004

Fingerprint

TATA Box
Mutagenesis
Retinoblastoma
Genes
Cell Line
Retinoblastoma Genes
Retinal Cone Photoreceptor Cells
Arrestin
RNA Polymerase II
Exons
arrestin3
Gene Expression
Proteins

Keywords

  • Cone photoreceptors
  • CRX, PCE-1
  • Y79 retinoblastoma

ASJC Scopus subject areas

  • Genetics

Cite this

Fujimaki, T., Huang, Z. Y., Kitagawa, H., Sakuma, H., Murakami, A., Kanai, A., ... Inana, G. (2004). Truncation and mutagenesis analysis of the human X-arrestin gene promoter. Gene, 339(1-2), 139-147. https://doi.org/10.1016/j.gene.2004.06.032

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 journalArticle

Fujimaki, T, Huang, ZY, Kitagawa, H, Sakuma, H, Murakami, A, Kanai, A, McLaren, MJ & Inana, G 2004, 'Truncation and mutagenesis analysis of the human X-arrestin gene promoter', Gene, vol. 339, no. 1-2, pp. 139-147. https://doi.org/10.1016/j.gene.2004.06.032
Fujimaki T, Huang ZY, Kitagawa H, Sakuma H, Murakami A, Kanai A et al. Truncation and mutagenesis analysis of the human X-arrestin gene promoter. Gene. 2004 Sep 15;339(1-2):139-147. https://doi.org/10.1016/j.gene.2004.06.032
Fujimaki, Takuro ; Huang, Zhen Yong ; Kitagawa, Hitoshi ; Sakuma, Hitoshi ; Murakami, Akira ; Kanai, Atsushi ; McLaren, Margaret J. ; Inana, George. / Truncation and mutagenesis analysis of the human X-arrestin gene promoter. In: Gene. 2004 ; Vol. 339, No. 1-2. pp. 139-147.
@article{4f9265ad13574bd5b759384c985d38de,
title = "Truncation and mutagenesis analysis of the human X-arrestin gene promoter",
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.",
keywords = "Cone photoreceptors, CRX, PCE-1, Y79 retinoblastoma",
author = "Takuro Fujimaki and Huang, {Zhen Yong} and Hitoshi Kitagawa and Hitoshi Sakuma and Akira Murakami and Atsushi Kanai and McLaren, {Margaret J.} and George Inana",
year = "2004",
month = "9",
day = "15",
doi = "10.1016/j.gene.2004.06.032",
language = "English",
volume = "339",
pages = "139--147",
journal = "Gene",
issn = "0378-1119",
publisher = "Elsevier",
number = "1-2",

}

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

VL - 339

SP - 139

EP - 147

JO - Gene

JF - Gene

SN - 0378-1119

IS - 1-2

ER -