Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P₂] is an important molecule for the progression of cytokinesis and accumulates the cleavage furrow during cytokinesis. Here, we investigated whether phospholipase C (PLC)-related catalytically inactive protein (PRIP), a metabolic modulator of PI(4,5)P₂, regulates PI(4,5)P₂-mediated cytokinesis. PRIP localized to the cleavage furrow during cytokinesis, and PRIP-knockdown HeLa cells displayed abnormal cytokinesis. Importantly, PI(4,5)P₂ accumulation at, and the localization of RhoA and phospho-myosin II regulatory light chain to, the cleavage furrow were reduced in the PRIP-knockdown cells. The overexpression of oculocerebrorenal syndrome of Lowe-1 (OCRL1), a phosphatidylinositol-5-phosphatase, in cells decreased PI(4,5)P₂ levels during early cytokinesis and showed cytokinesis abnormalities. However, these abnormal cytokinesis phenotypes were ameliorated by the co-expression of PRIP but not by a PI(4,5)P₂-unbound PRIP mutant. Collectively, PRIP is a component at the cleavage furrow to maintain PI(4,5)P₂ metabolism and regulates RhoA-dependent progression of cytokinesis.