ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR251/025/SRR25184425/SRR25184425_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR251/026/SRR25184426/SRR25184426_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR251/027/SRR25184427/SRR25184427_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR251/028/SRR25184428/SRR25184428_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR251/026/SRR25184426/SRR25184426_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR251/025/SRR25184425/SRR25184425_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR251/027/SRR25184427/SRR25184427_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR251/028/SRR25184428/SRR25184428_2.fastq.gzprimaryOK200GenomicsUniversity of Texas at Austinhttps://www.ebi.ac.uk/ena/browser/view/PRJNA992449Flotillin-1 contributes to invasion and metastasis in triple negative breast cancer (TNBC). Palmitoylation, the process of conjugating palmitoyl-CoA to proteins, plays an essential role in protein stability and trafficking. Flotillin-1 is modified by palmitoylation, however, the role of its palmitoylation in the context of metastasis has not been explored. Using palmitoylation defective flotillin-1 constructs, we have demonstrated that flotillin-1 palmitoylation contributes to its stability and metastatic capabilities in vivo. Further investigation led to the identification of zDHHC5 as the main palmitoyl acyl transferase responsible for palmitoylating flotillin-1, which also contributed to its stability by preventing its poly-ubiquitylation. To assess the ability to target flotillin-1 palmitoylation therapeutically, we designed a competitive peptide, which displayed efficacy in blocking flotillin-1 palmitoylation in vitro without altering palmitoylation of other zDHHC5 substrates, highlighting its specificity. Additionally, multiple TNBC tumor models expressing a doxycycline inducible flotillin-1 palmitoylation inhibiting peptide construct displayed attenuated tumor growth and lung metastasis. The current study has demonstrated the palmitoylation of flotillin-1, a known metastasis inducing protein, to be essential for its protein stability. The demonstrated methods in blocking its palmitoylation through the delivery of a competitive peptide provide proof-of-concept data for further development as a potential targeted therapeutic in TNBC.ENAfalseFlotillin-1 palmitoylation is essential for its stability and subsequent tumor promoting capabilities.2024-03-302024-03-30PRJNA992449