We originally cloned SAFB1 as a transcriptional repressor, and it has since been implicated in a number of cellular processes including stress response, apoptosis, RNA processing, hormone response, immortalization, and transformation. During the last funding period we performed detailed structure function studies which identified a transcriptional repression domain, we found that sumoylation of SAFB1 was necessary for its co-repressor activity, and finally, we generated and characterized SAFB1-null mice which showed prenatal and neonatal lethality with surviving mice showing dramatic defects in the development and function of the reproductive system. To gain more insight into SAFB1's role in ERalpha action, we performed an unbiased screen using SAFB1 siRNA knockdown followed by microarray analysis. This experiment revealed a significant role for SAFB1 in estrogen-mediated repression of gene expression, and candidate genes that require SAFB1 for estrogen-mediated downregulation were identified. Here we propose to study how SAFB1 mediates estrogen repression of gene expression. Specifically, we will i) determine how SAFB1 SAFB1 mediates transcriptional repression of estrogen-responsive target genes, ii) characterize HDAC7 as a critical player in SAFB1-mediated repression of estrogen regulated genes, and iii) determine how posttranslational modification by sumoylation affects SAFB1's co-repressor function. This work is highly significant since it provides studies of fundamental mechanisms which may contribute to a change in paradigm of ERalpha's action. While ERalpha has mainly been studied as a transcriptional activator, the study of estrogen- mediated repression of gene expression, and in particular the role of co-repressors, is an understudied area. 1

Public Health Relevance

SAFB1 is a transcriptional co-repressor, and it has since been implicated in a number of cellular processes including stress response, apoptosis, RNA processing, hormone response, immortalization, and transformation. We have discovered that SAFB1 plays a significant role in estrogen-mediated repression of gene expression, which is an understudied yet critical process in estrogen receptor action. Here we propose to study the in-depth mechanism of this repression, including its action on promoter and enhancer region of candidate gene, the role of its interacting proteins, and its posttranslational modification by sumoylation.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA097213-10
Application #
8311847
Study Section
Molecular and Cellular Endocrinology Study Section (MCE)
Program Officer
Sathyamoorthy, Neeraja
Project Start
2002-08-01
Project End
2014-08-31
Budget Start
2012-09-01
Budget End
2014-08-31
Support Year
10
Fiscal Year
2012
Total Cost
$293,910
Indirect Cost
$99,910
Name
University of Pittsburgh
Department
Pharmacology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Jiang, Shiming; Katz, Tiffany A; Garee, Jason P et al. (2015) Scaffold attachment factor B2 (SAFB2)-null mice reveal non-redundant functions of SAFB2 compared with its paralog, SAFB1. Dis Model Mech 8:1121-7
Nayak, Shweta R; Harrington, Emily; Boone, David et al. (2015) A Role for Histone H2B Variants in Endocrine-Resistant Breast Cancer. Horm Cancer 6:214-24
Mukhopadhyay, N K; Kim, J; You, S et al. (2014) Scaffold attachment factor B1 regulates the androgen receptor in concert with the growth inhibitory kinase MST1 and the methyltransferase EZH2. Oncogene 33:3235-45
Pathiraja, Thushangi N; Nayak, Shweta R; Xi, Yuanxin et al. (2014) Epigenetic reprogramming of HOXC10 in endocrine-resistant breast cancer. Sci Transl Med 6:229ra41
Hernandez-Hernandez, J Manuel; Mallappa, Chandrashekara; Nasipak, Brian T et al. (2013) The Scaffold attachment factor b1 (Safb1) regulates myogenic differentiation by facilitating the transition of myogenic gene chromatin from a repressed to an activated state. Nucleic Acids Res 41:5704-16
Oesterreich, Steffi; Brufsky, Adam M; Davidson, Nancy E (2013) Using mice to treat (wo)men: mining genetic changes in patient xenografts to attack breast cancer. Cell Rep 4:1061-2
Casa, Angelo J; Potter, Adam S; Malik, Simeen et al. (2012) Estrogen and insulin-like growth factor-I (IGF-I) independently down-regulate critical repressors of breast cancer growth. Breast Cancer Res Treat 132:61-73
Garee, Jason P; Meyer, Rene; Oesterreich, Steffi (2011) Co-repressor activity of scaffold attachment factor B1 requires sumoylation. Biochem Biophys Res Commun 408:516-22
Malik, Simeen; Jiang, Shiming; Garee, Jason P et al. (2010) Histone deacetylase 7 and FoxA1 in estrogen-mediated repression of RPRM. Mol Cell Biol 30:399-412
Garee, Jason P; Oesterreich, Steffi (2010) SAFB1's multiple functions in biological control-lots still to be done! J Cell Biochem 109:312-9

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