Syntaxin 1A Gene Is Negatively Regulated in a Cell/Tissue Specific Manner by YY1 Transcription Factor, Which Binds to the −183 to −137 Promoter Region Together with Gene Silencing Factors Including Histone Deacetylase

The HPC-1/syntaxin 1A (<i>Stx1a</i>) gene, which is involved in synaptic transmission and neurodevelopmental disorders, is a TATA-less gene with several transcription start sites. It is activated by the binding of Sp1 and acetylated histone H3 to the −204 to +2 core promoter region (CPR) in neuronal cell/tissue. Furthermore, it is depressed by the association of class 1 histone deacetylases (HDACs) to <i>Stx1a</i>–CPR in non-neuronal cell/tissue. To further clarify the factors characterizing <i>Stx1a</i> gene silencing in non-neuronal cell/tissue not expressing <i>Stx1a</i>, we attempted to identify the promoter region forming DNA–protein complex only in non-neuronal cells. Electrophoresis mobility shift assays (EMSA) demonstrated that the −183 to −137 OL2 promoter region forms DNA–protein complex only in non-neuronal fetal rat skin keratinocyte (FRSK) cells which do not express <i>Stx1a</i>. Furthermore, the Yin-Yang 1 (YY1) transcription factor binds to the −183 to −137 promoter region of <i>Stx1a</i> in FRSK cells, as shown by competitive EMSA and supershift assay. Chromatin immunoprecipitation assay revealed that YY1 in vivo associates to <i>Stx1a</i>–CPR in cell/tissue not expressing <i>Stx1a</i> and that trichostatin A treatment in FRSK cells decreases the high-level association of YY1 to <i>Stx1a</i>-CPR in default. Reporter assay indicated that YY1 negatively regulates <i>Stx1a</i> transcription. Finally, mass spectrometry analysis showed that gene silencing factors, including HDAC1, associate onto the −183 to −137 promoter region together with YY1. The current study is the first to report that <i>Stx1a</i> transcription is negatively regulated in a cell/tissue-specific manner by YY1 transcription factor, which binds to the −183 to −137 promoter region together with gene silencing factors, including HDAC.