Mutational analysis of the C-terminal FATC domain of Saccharomyces cerevisiae Tra1.

Current Genetics
Stephen M T HokeChristopher J Brandl

Abstract

Tra1 is a component of the Saccharomyces cerevisiae SAGA and NuA4 complexes and a member of the PIKK family, which contain a C-terminal phosphatidylinositol 3-kinase-like (PI3K) domain followed by a 35-residue FATC domain. Single residue changes of L3733A and F3744A, within the FATC domain, resulted in transcriptional changes and phenotypes that were similar but not identical to those caused by mutations in the PI3K domain or deletions of other SAGA or NuA4 components. The distinct nature of the FATC mutations was also apparent from the additive effect of tra1-L3733A with SAGA, NuA4, and tra1 PI3K domain mutations. Tra1-L3733A associates with SAGA and NuA4 components and with the Gal4 activation domain, to the same extent as wild-type Tra1; however, steady-state levels of Tra1-L3733A were reduced. We suggest that decreased stability of Tra1-L3733A accounts for the phenotypes since intragenic suppressors of tra1-L3733A restored Tra1 levels, and reducing wild-type Tra1 led to comparable growth defects. Also supporting a key role for the FATC domain in the structure/function of Tra1, addition of a C-terminal glycine residue resulted in decreased association with Spt7 and Esa1, and loss of cellular viability. These findings demonst...Continue Reading

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Citations

Mar 24, 2012·BMC Genetics·Stephanie KvasChristopher J Brandl
Nov 27, 2019·Biochemical Society Transactions·Alberto Elías-VillalobosDominique Helmlinger
Aug 13, 2013·G3 : Genes - Genomes - Genetics·Lance F DaSilvaChristopher J Brandl
Mar 22, 2018·Nature Communications·Xuejuan WangGang Cai
May 14, 2016·G3 : Genes - Genomes - Genetics·Kyle S HoffmanChristopher J Brandl

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Datasets Mentioned

BETA
GSE18591

Methods Mentioned

BETA
acetylation
PCR
dot blots
immunoprecipitation
dot
immunoprecipitations
tandem affinity purification
affinity purification
two-hybrid

Software Mentioned

Rosetta Resolver
novoindex
ClustalW
AlphaImager 3400
AlphaImager
novoalign
CLUSTER

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