Dysfunctional CAF-I reveals its role in cell cycle progression and differential regulation of gene silencing. Issue 22 (17th November 2019)

• Record Type: Journal Article
• Authors: Rowlands, Hollie; Shaban, Kholoud; Cheng, Ashley; Foster, Barret; Yankulov, Krassimir
• Is Part Of: Cell cycle. Volume 18:Issue 22(2019)
• Page Start: 3223
• Page End: 3236
• Publication Date: 2019-11-17
• Subjects: Chromatin assembly factor I -- cyclin dependent kinase (CDK) -- Dbf4-dependent kinase (DDK) -- cell cycle -- flocculation -- FLO genes; Cell cycle -- Periodicals; 571.84377
• DOI: 10.1080/15384101.2019.1673100
• Languages: English
• ISSNs: 1538-4101

Abstract

ABSTRACT: Chromatin Assembly Factor I (CAF-I) plays a central role in the reassembly of H3/H4 histones during DNA replication. In S. cerevisiae CAF-I is not essential and its loss is associated with reduced gene silencing at telomeres and increased sensitivity to DNA damage. Two kinases, Cyclin Dependent Kinase (CDK) and Dbf4-Dependent Kinase (DDK), are known to phosphorylate the Cac1p subunit of CAF-I, but their role in the regulation of CAF-I activity is not well understood. In this study we systematically mutated the phosphorylation target sites of these kinases. We show that concomitant mutations of the CDK and DDK target sites of Cac1p lead to growth retardation and significant cell cycle defects, altered cell morphology and increased sensitivity to DNA damage. Surprisingly, some mutations also produced flocculation, a phenotype that is lost in most laboratory strains, and displayed elevated expression of FLO genes. None of these effects is observed upon the destruction of CAF-I. In contrast, the mutations that caused flocculation did not affect gene silencing at the mating type and subtelomeric loci. We conclude that dysfunctional CAF-I produces severe phenotypes, which reveal a possible role of CAF-I in the coordination of DNA replication, chromatin reassembly and cell cycle progression. Our study highlights the role of phosphorylation of Cac1p by CDK and a putative role for DDK in the transmission and re-assembly of chromatin during DNA replication.