Role of the CCAAT-binding protein NFY in SCA17 pathogenesis

Li Ching Lee, Chiung Mei Chen, Hao Chun Wang, Hsiao Han Hsieh, I. Sheng Chiu, Ming Tsan Su, Hsiu Mei Hsieh-Li, Chung Hsin Wu, Guan Chiun Lee, Guey Jen Lee-Chen, Jung Yaw Lin

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Spinocerebellar ataxia 17 (SCA17) is caused by expansion of the polyglutamine (polyQ) tract in human TATA-box binding protein (TBP) that is ubiquitously expressed in both central nervous system and peripheral tissues. The spectrum of SCA17 clinical presentation is broad. The precise pathogenic mechanism in SCA17 remains unclear. Previously proteomics study using a cellular model of SCA17 has revealed reduced expression of heat shock 70 kDa protein 5 (HSPA5) and heat shock 70 kDa protein 8 (HSPA8), suggesting that impaired protein folding may contribute to the cell dysfunction of SCA17 (Lee et al., 2009). In lymphoblastoid cells, HSPA5 and HSPA8 expression levels in cells with mutant TBP were also significantly lower than that of the control cells (Chen et al., 2010). As nuclear transcription factor Y (NFY) has been reported to regulate HSPA5 transcription, we focused on if NFY activity and HSPA5 expression in SCA17 cells are altered. Here, we show that TBP interacts with NFY subunit A (NFYA) in HEK-293 cells and NFYA incorporated into mutant TBP aggregates. In both HEK-293 and SH-SY5Y cells expressing TBP/Q61~79, the level of soluble NFYA was significantly reduced. In vitro binding assay revealed that the interaction between TBP and NFYA is direct. HSPA5 luciferase reporter assay and endogenous HSPA5 expression analysis in NFYA cDNA and siRNA transfection cells further clarified the important role of NFYA in regulating HSPA5 transcription. In SCA17 cells, HSPA5 promoter activity was activated as a compensatory response before aggregate formation. NFYA dysfunction was indicated in SCA17 cells as HSPA5 promoter activity reduced along with TBP aggregate formation. Because essential roles of HSPA5 in protection from neuronal apoptosis have been shown in a mouse model, NFYA could be a target of mutant TBP in SCA17.

Original languageEnglish
Article numbere35302
JournalPloS one
Volume7
Issue number4
DOIs
Publication statusPublished - 2012 Apr 17

Fingerprint

TATA-Box Binding Protein
TATA box
binding proteins
Carrier Proteins
Transcription Factors
pathogenesis
transcription factors
cells
Mutant Proteins
HSP70 Heat-Shock Proteins
protein aggregates
Transcription
Assays
Protein folding
mutants
Neurology
transcription (genetics)
promoter regions
Spinocerebellar Ataxia 17
nuclear factor Y

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Lee, L. C., Chen, C. M., Wang, H. C., Hsieh, H. H., Chiu, I. S., Su, M. T., ... Lin, J. Y. (2012). Role of the CCAAT-binding protein NFY in SCA17 pathogenesis. PloS one, 7(4), [e35302]. https://doi.org/10.1371/journal.pone.0035302

Role of the CCAAT-binding protein NFY in SCA17 pathogenesis. / Lee, Li Ching; Chen, Chiung Mei; Wang, Hao Chun; Hsieh, Hsiao Han; Chiu, I. Sheng; Su, Ming Tsan; Hsieh-Li, Hsiu Mei; Wu, Chung Hsin; Lee, Guan Chiun; Lee-Chen, Guey Jen; Lin, Jung Yaw.

In: PloS one, Vol. 7, No. 4, e35302, 17.04.2012.

Research output: Contribution to journalArticle

Lee LC, Chen CM, Wang HC, Hsieh HH, Chiu IS, Su MT et al. Role of the CCAAT-binding protein NFY in SCA17 pathogenesis. PloS one. 2012 Apr 17;7(4). e35302. https://doi.org/10.1371/journal.pone.0035302
Lee, Li Ching ; Chen, Chiung Mei ; Wang, Hao Chun ; Hsieh, Hsiao Han ; Chiu, I. Sheng ; Su, Ming Tsan ; Hsieh-Li, Hsiu Mei ; Wu, Chung Hsin ; Lee, Guan Chiun ; Lee-Chen, Guey Jen ; Lin, Jung Yaw. / Role of the CCAAT-binding protein NFY in SCA17 pathogenesis. In: PloS one. 2012 ; Vol. 7, No. 4.
@article{daf4cb7706f446ecacd535f61d0b0401,
title = "Role of the CCAAT-binding protein NFY in SCA17 pathogenesis",
abstract = "Spinocerebellar ataxia 17 (SCA17) is caused by expansion of the polyglutamine (polyQ) tract in human TATA-box binding protein (TBP) that is ubiquitously expressed in both central nervous system and peripheral tissues. The spectrum of SCA17 clinical presentation is broad. The precise pathogenic mechanism in SCA17 remains unclear. Previously proteomics study using a cellular model of SCA17 has revealed reduced expression of heat shock 70 kDa protein 5 (HSPA5) and heat shock 70 kDa protein 8 (HSPA8), suggesting that impaired protein folding may contribute to the cell dysfunction of SCA17 (Lee et al., 2009). In lymphoblastoid cells, HSPA5 and HSPA8 expression levels in cells with mutant TBP were also significantly lower than that of the control cells (Chen et al., 2010). As nuclear transcription factor Y (NFY) has been reported to regulate HSPA5 transcription, we focused on if NFY activity and HSPA5 expression in SCA17 cells are altered. Here, we show that TBP interacts with NFY subunit A (NFYA) in HEK-293 cells and NFYA incorporated into mutant TBP aggregates. In both HEK-293 and SH-SY5Y cells expressing TBP/Q61~79, the level of soluble NFYA was significantly reduced. In vitro binding assay revealed that the interaction between TBP and NFYA is direct. HSPA5 luciferase reporter assay and endogenous HSPA5 expression analysis in NFYA cDNA and siRNA transfection cells further clarified the important role of NFYA in regulating HSPA5 transcription. In SCA17 cells, HSPA5 promoter activity was activated as a compensatory response before aggregate formation. NFYA dysfunction was indicated in SCA17 cells as HSPA5 promoter activity reduced along with TBP aggregate formation. Because essential roles of HSPA5 in protection from neuronal apoptosis have been shown in a mouse model, NFYA could be a target of mutant TBP in SCA17.",
author = "Lee, {Li Ching} and Chen, {Chiung Mei} and Wang, {Hao Chun} and Hsieh, {Hsiao Han} and Chiu, {I. Sheng} and Su, {Ming Tsan} and Hsieh-Li, {Hsiu Mei} and Wu, {Chung Hsin} and Lee, {Guan Chiun} and Lee-Chen, {Guey Jen} and Lin, {Jung Yaw}",
year = "2012",
month = "4",
day = "17",
doi = "10.1371/journal.pone.0035302",
language = "English",
volume = "7",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "4",

}

TY - JOUR

T1 - Role of the CCAAT-binding protein NFY in SCA17 pathogenesis

AU - Lee, Li Ching

AU - Chen, Chiung Mei

AU - Wang, Hao Chun

AU - Hsieh, Hsiao Han

AU - Chiu, I. Sheng

AU - Su, Ming Tsan

AU - Hsieh-Li, Hsiu Mei

AU - Wu, Chung Hsin

AU - Lee, Guan Chiun

AU - Lee-Chen, Guey Jen

AU - Lin, Jung Yaw

PY - 2012/4/17

Y1 - 2012/4/17

N2 - Spinocerebellar ataxia 17 (SCA17) is caused by expansion of the polyglutamine (polyQ) tract in human TATA-box binding protein (TBP) that is ubiquitously expressed in both central nervous system and peripheral tissues. The spectrum of SCA17 clinical presentation is broad. The precise pathogenic mechanism in SCA17 remains unclear. Previously proteomics study using a cellular model of SCA17 has revealed reduced expression of heat shock 70 kDa protein 5 (HSPA5) and heat shock 70 kDa protein 8 (HSPA8), suggesting that impaired protein folding may contribute to the cell dysfunction of SCA17 (Lee et al., 2009). In lymphoblastoid cells, HSPA5 and HSPA8 expression levels in cells with mutant TBP were also significantly lower than that of the control cells (Chen et al., 2010). As nuclear transcription factor Y (NFY) has been reported to regulate HSPA5 transcription, we focused on if NFY activity and HSPA5 expression in SCA17 cells are altered. Here, we show that TBP interacts with NFY subunit A (NFYA) in HEK-293 cells and NFYA incorporated into mutant TBP aggregates. In both HEK-293 and SH-SY5Y cells expressing TBP/Q61~79, the level of soluble NFYA was significantly reduced. In vitro binding assay revealed that the interaction between TBP and NFYA is direct. HSPA5 luciferase reporter assay and endogenous HSPA5 expression analysis in NFYA cDNA and siRNA transfection cells further clarified the important role of NFYA in regulating HSPA5 transcription. In SCA17 cells, HSPA5 promoter activity was activated as a compensatory response before aggregate formation. NFYA dysfunction was indicated in SCA17 cells as HSPA5 promoter activity reduced along with TBP aggregate formation. Because essential roles of HSPA5 in protection from neuronal apoptosis have been shown in a mouse model, NFYA could be a target of mutant TBP in SCA17.

AB - Spinocerebellar ataxia 17 (SCA17) is caused by expansion of the polyglutamine (polyQ) tract in human TATA-box binding protein (TBP) that is ubiquitously expressed in both central nervous system and peripheral tissues. The spectrum of SCA17 clinical presentation is broad. The precise pathogenic mechanism in SCA17 remains unclear. Previously proteomics study using a cellular model of SCA17 has revealed reduced expression of heat shock 70 kDa protein 5 (HSPA5) and heat shock 70 kDa protein 8 (HSPA8), suggesting that impaired protein folding may contribute to the cell dysfunction of SCA17 (Lee et al., 2009). In lymphoblastoid cells, HSPA5 and HSPA8 expression levels in cells with mutant TBP were also significantly lower than that of the control cells (Chen et al., 2010). As nuclear transcription factor Y (NFY) has been reported to regulate HSPA5 transcription, we focused on if NFY activity and HSPA5 expression in SCA17 cells are altered. Here, we show that TBP interacts with NFY subunit A (NFYA) in HEK-293 cells and NFYA incorporated into mutant TBP aggregates. In both HEK-293 and SH-SY5Y cells expressing TBP/Q61~79, the level of soluble NFYA was significantly reduced. In vitro binding assay revealed that the interaction between TBP and NFYA is direct. HSPA5 luciferase reporter assay and endogenous HSPA5 expression analysis in NFYA cDNA and siRNA transfection cells further clarified the important role of NFYA in regulating HSPA5 transcription. In SCA17 cells, HSPA5 promoter activity was activated as a compensatory response before aggregate formation. NFYA dysfunction was indicated in SCA17 cells as HSPA5 promoter activity reduced along with TBP aggregate formation. Because essential roles of HSPA5 in protection from neuronal apoptosis have been shown in a mouse model, NFYA could be a target of mutant TBP in SCA17.

UR - http://www.scopus.com/inward/record.url?scp=84859861243&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84859861243&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0035302

DO - 10.1371/journal.pone.0035302

M3 - Article

C2 - 22530004

AN - SCOPUS:84859861243

VL - 7

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 4

M1 - e35302

ER -