Analysis of transcriptional modulation of the presenilin 1 gene promoter by ZNF237, a candidate binding partner of the Ets transcription factor ERM

Abstract

DNA sequences required for the expression of the human presenilin 1 (PS1) gene have been identified between -118 and +178 flanking the major initiation site (+1) mapped in SK-N-SH cells. Several Ets sites are located both upstream as well as downstream from the +1 site, including an Ets motif present at -10 that controls 90% of transcription in SK-N-SH cells. However in SH-SY5Y cells transcription initiates further downstream and requires an alternative set of promoter elements including a +90 Ets motif. Ets2, ER81, ERM and Elk1 were identified by yeast one-hybrid selection in a human brain cDNA library using the -10 Ets motif as a bait. We have shown that ERM recognizes specifically Ets motifs on the PS1 promoter located at -10 as well as downstream at +90, +129 and +165 and activates PS1 transcription with promoter fragments whether or not they contain the -10 Ets site. We have now searched for ERM interacting proteins by yeast two-hybrid selection in a human brain cDNA library using the C-terminal 415 amino acid of ERM as a bait. One of the interacting proteins was ZNF237, a member of the MYM gene family. It is widely expressed in different tissues in eukaryotes under several forms derived by alternative splicing, including a large 382 amino acid form containing a single MYM domain, and 2 shorter forms of 208 and 213 amino acids respectively that do not. We show that both the 382 as well as the 208 amino acid forms are expressed in SK-N-SH cells but not in SH-SY5Y cells. Both forms interact with ERM and repress the transcription of PS1 in SH-SY5Y cells. The effect of both C-terminal and N-terminal deletions indicate that the Nterminal 120 amino acid region is required for interaction with ERM in yeast and furthermore single amino acid mutations show that residues 112 and 114 play an important role. The repression of transcription in SH-SY5Y cells also appears to require the N-terminal potion of ZNF237 and was affected by mutation of the amino acid 112. Data from electrophoretic mobility shift assays indicate that ERM and possibly ZNF237, interact with a fragment of the PS1 promoter.

Repression of transcription of presenilin‐1 inhibits γ‐secretase independent ER Ca2+ leak that is impaired by FAD mutations

“…We have previously shown that ZNF237 and CHD3 repress PS1 transcription and PS1 protein level in human neuroblastoma cells (Lee and Das 2010; Pastorcic and Das 2007a,b). To determine the role of ZNF237 and CHD3 in controlling the function of ER Ca 2+ leak channels, we performed Ca 2+ ‐imaging experiments in the presence of extracellular Ca 2+ in SK‐N‐SH cells which were transiently transfected with pCMV‐Tag2 or pCMV‐Tag2‐ZNF237 or pCMV‐Tag2‐CHD3 constructs for 48 h. Cell extracts were prepared and levels of FLAG‐ZNF237 and FLAG‐CHD3 were determined using western blot analysis using anti‐FLAG antibody, and PS1 level was determined by anti‐PS1 antibody as described before (Pastorcic and Das 2007b; Lee and Das 2010).…”

“…Wild type PS1 cDNA was cloned into pCDNA3 vector to produce pCDNA3.PS1. Constructions of pCMV.p53, pCMV‐Tag2‐ZNF237, pCMV‐Tag2‐CHD3 were reported previously (Pastorcic and Das 2000, 2007b,a). SK‐N‐SH cells were seeded in culture dishes on to glass coverslips the day before transfection.…”

“…We have shown that zinc finger protein ZNF237, and chromodomain helicase DNA‐binding protein (CHD3) also down‐regulate PS1 transcription (Pastorcic and Das 2007b,a; Lee and Das 2010). In this report, we show that suppression of PS1 transcription by JNK inhibitor SP600125, p53, ZNF237, and CHD3 also inhibit PS1‐facilitated ER Ca 2+ leak channels resulting in Ca 2+ signaling defects in human neuroblastoma SK‐N‐SH cells.…”

Application of array comparative genomic hybridization in 256 patients with developmental delay or intellectual disability

“…Functionally, PSPC1 was proposed to participate in the regulation of transcription (Passon et al 2011). ZMYM5 belongs to the zinc finger MYM-type family and probably has molecular functions, such as metal ion binding or zinc ion binding (Pastorcic and Das 2007). Although the phenotype–genotype correlation remains unclear and one smaller deletion within this region was reported in the control group (DGV; nsv455826) (Itsara et al 2009), we propose that the identified de novo 13q12.11 deletion could contribute to the clinical features observed in our patient.

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Transcriptional regulation of the presenilin-1 gene controls gamma-secretase activity

“…Therefore, we have studied the regulation of transcription of the human PS1 gene. We have shown that Ets1/2 transcription factors upregulate PS1 transcription (14)(15)(16)(17), whereas transcription factors p53, ZNF237, and CHD3 downregulate PS1 transcription (15,18,19). We have recently shown that inhibition of basal JNK activity by JNK inhibitor SP600125 represses PS1 transcription by a p53 dependent mechanism and also reduces PS1 protein level (1).…”