Septin-associated protein kinase Gin4 affects localization and phosphorylation of Chs4, the regulatory subunit of the Baker's yeast chitin synthase III complex

Abstract

Chitin is mainly formed by the chitin synthase III complex (CSIII) in yeast cells. This complex is considered to be composed of the catalytic subunit Chs3 and the regulatory subunit Chs4, both of which are phosphoproteins and transported to the plasma membrane by different trafficking routes. During cytokinesis, Chs3 associates with Chs4 and other proteins at the septin ring, which results in an active CSIII complex. In this study, we focused on the role of Chs4 as a regulatory subunit of the CSIII complex. We analyzed the dynamic localization and interaction of Chs3 and Chs4 during cell division, and found that both proteins transiently co-localize and physically interact only during bud formation and later in a period during septum formation and cytokinesis. To identify unknown binding partners of Chs4, we conducted different screening approaches, which yielded several novel candidates of Chs4-binding proteins including the septin-associated kinase Gin4. Our further studies confirmed this interaction and provided first evidence that Chs4 phosphorylation is partially dependent on Gin4, which is required for proper localization of Chs4 at the bud neck.