KCTD Proteins Have Redundant Functions in Controlling Cellular Growth

• Published in: International journal of molecular sciences Vol. 25; no. 9; p. 4993
• Main Authors: Rizk, Robert, Devost, Dominic, Pétrin, Darlaine, Hébert, Terence E
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.05.2024
• Subjects: Apoptosis; Cell cycle; Cell growth; Cell Proliferation - genetics; cellular signaling; Cloning; CRISPR; CRISPR-Cas Systems; Ethylenediaminetetraacetic acid; G proteins; Gene Editing; Gene expression; Gene Expression Regulation; Gene Knockout Techniques; Genetic transcription; Genome editing; Genomes; Genomics; Growth; HEK293 Cells; Heterotrimeric G proteins; Humans; KCTD proteins; Kinases; Potassium Channels - genetics; Potassium Channels - metabolism; Protein Isoforms - genetics; Protein Isoforms - metabolism; Proteins; RNA polymerase; cell growth; cellular signaling; KCTD proteins; Heterotrimeric G proteins
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25094993

We explored the functional redundancy of three structurally related KCTD (Potassium Channel Tetramerization Domain) proteins, KCTD2, KCTD5, and KCTD17, by progressively knocking them out in HEK 293 cells using CRISPR/Cas9 genome editing. After validating the knockout, we assessed the effects of progressive knockout on cell growth and gene expression. We noted that the progressive effects of knockout of KCTD isoforms on cell growth were most pervasive when all three isoforms were deleted, suggesting some functions were conserved between them. This was also reflected in progressive changes in gene expression. Our previous work indicated that Gβ1 was involved in the transcriptional control of gene expression, so we compared the gene expression patterns between GNB1 and KCTD KO. Knockout of GNB1 led to numerous changes in the expression levels of other G protein subunit genes, while knockout of KCTD isoforms had the opposite effect, presumably because of their role in regulating levels of Gβ1. Our work demonstrates a unique relationship between KCTD proteins and Gβ1 and a global role for this subfamily of KCTD proteins in maintaining the ability of cells to survive and proliferate.