Identification of druggable targets from the interactome of the Androgen Receptor and Serum Response Factor pathways in prostate cancer

Background: The Androgen Receptor (AR) pathway is crucial in driving the progression of prostate cancer (PCa) to an advanced state. Despite the introduction of second-generation AR antagonists, such as enzalutamide, majority of patients develop resistance. Several mechanisms of resistance have been identified, including the constitutive activation of the AR pathway, the emergence of AR spliced variants, and the influence of other signalling pathways. The Serum Response Factor (SRF) was previously identified as a possible player of resistance involved in a crosstalk with the AR signalling pathway. Elevated SRF levels in PCa patients were associated with disease progression and resistance to enzalutamide. However, the molecular mediators of the crosstalk between SRF and AR still need to be elucidated. The objective of this study was to identify common interactors of the AR/SRF crosstalk as therapeutic targets.
Methods: Here we used affinity purification mass spectrometry (MS) following im..., Co-Immunoprecipitation Assay
Prior to cellular lysis, the antibody solution was prepared adding 2μg of either SRF (Novus, Biotechne), AR (Santa Cruz, California, US), Ms IgG or Rb IgG to 20μL A/G protein beads (Pierce) and 300μL PBS (Gibco). The beads/antibody mixture was incubated for an hour on a rotator at 4°C. Following incubation, the mixture was washed with ice-cold lysis buffer 3 times.  Cells were scraped with 500μL of lysis buffer (500μL of 1% Triton x100, 1mL of 20mM Tris-HCl pH7.5, 1.5mL of 150mM NaCl and 50μL of 1mM MgCl2) and incubated for 10 mins on ice. 1mg of protein was added to the beads/antibody mix and incubated for 1 hour on a rotator at 4°C. Samples were washed 3 times in ice-cold lysis buffer.
Peptide elution and digestion
Following Co-IP, the peptides in each sample were eluted with 60μL of ice-cold Elution Buffer I (0.012g Urea, 50μL of 1M Tris-HCl pH7.5) and 5μg/mL Trypsin (Promega, Seq Grade Modified) for 30 mins at RT. Samples were then centrifuged at 3000rpm..., , # List of peptides precipitated with AR and SRF

[https://doi.org/10.5061/dryad.63xsj3vbb](https://doi.org/10.5061/dryad.63xsj3vbb)

## Description of the data and file structure

This dataset was created to identify common interactors of the androgen receptor (AR) and serum response factor (SRF) in prostate cancer. AR and SRF were precipitated using specific antibodies followed by affinity purification mass spectrometry.

### Files and variables

#### File: 220209kw\_HaleemaIP\_1\_S1-A2\_1\_8007.d\_Caitriona\_Scaife.zip

**Description:** AR co-IP of ARsiRNA N1 mass spec row data

#### File: 220209kw\_HaleemaIP\_2\_S1-A3\_1\_8008.d\_Caitriona\_Scaife.zip

**Description:** AR co-IP of endogenous AR N1 mass spec row data

#### File: 220209kw\_HaleemaIP\_3\_S1-A4\_1\_8009.d\_Caitriona\_Scaife.zip

**Description:** AR co-IP of endogenous AR post DHT stimulation N1 mass spec row data

#### File: 220209kw\_HaleemaIP\_4\_S1-A5\_1\_8010.d\_Caitriona\_Scaife.zip

**Description:** AR co-IP of ARs...