biostudies-literatureGao GShanghai Municipal Science and Technology CommissionNational Natural Science Foundation of ChinaNational Key Research and Development Program of Chinae80865https://www.ebi.ac.uk/biostudies/studies/S-EPMC9714970biostudies-literatureUnknown11Unbiased genetic screens implicated a number of uncharacterized genes in hearing loss, suggesting some biological processes required for auditory function remain unexplored. Loss of <i>Kiaa1024L</i>/<i>Minar2</i>, a previously understudied gene, caused deafness in mice, but how it functioned in the hearing was unclear. Here, we show that disruption of <i>kiaa1024L/minar2</i> causes hearing loss in the zebrafish. Defects in mechanotransduction, longer and thinner hair bundles, and enlarged apical lysosomes in hair cells are observed in the <i>kiaa1024L/minar2</i> mutant. In cultured cells, Kiaa1024L/Minar2 is mainly localized to lysosomes, and its overexpression recruits cholesterol and increases cholesterol labeling. Strikingly, cholesterol is highly enriched in the hair bundle membrane, and loss of <i>kiaa1024L/minar2</i> reduces cholesterol localization to the hair bundles. Lowering cholesterol levels aggravates, while increasing cholesterol levels rescues the hair cell defects in the <i>kiaa1024L/minar2</i> mutant. Therefore, cholesterol plays an essential role in hair bundles, and Kiaa1024L/Minar2 regulates cholesterol distribution and homeostasis to ensure normal hearing.eLifeKiaa1024L/Minar2 is essential for hearing by regulating cholesterol distribution in hair bundles.PMC97149702018YFA080100031571067Major Project No.2018SHZDZX01Peng GZhang HGao GGuo SZhang CZhang QfalseKiaa1024L/Minar2 is essential for hearing by regulating cholesterol distribution in hair bundles.Unbiased genetic screens implicated a number of uncharacterized genes in hearing loss, suggesting some biological processes required for auditory function remain unexplored. Loss of <i>Kiaa1024L</i>/<i>Minar2</i>, a previously understudied gene, caused deafness in mice, but how it functioned in the hearing was unclear. Here, we show that disruption of <i>kiaa1024L/minar2</i> causes hearing loss in the zebrafish. Defects in mechanotransduction, longer and thinner hair bundles, and enlarged apical lysosomes in hair cells are observed in the <i>kiaa1024L/minar2</i> mutant. In cultured cells, Kiaa1024L/Minar2 is mainly localized to lysosomes, and its overexpression recruits cholesterol and increases cholesterol labeling. Strikingly, cholesterol is highly enriched in the hair bundle membrane, and loss of <i>kiaa1024L/minar2</i> reduces cholesterol localization to the hair bundles. Lowering cholesterol levels aggravates, while increasing cholesterol levels rescues the hair cell defects in the <i>kiaa1024L/minar2</i> mutant. Therefore, cholesterol plays an essential role in hair bundles, and Kiaa1024L/Minar2 regulates cholesterol distribution and homeostasis to ensure normal hearing.2022-01-01T00:00:00Z2022 Nov2025-04-19T03:31:49.693Z2025-04-07T13:41:42.302ZS-EPMC97149703631796210.7554/eLife.80865