Saccharomyces cerevisiae Phospholipase C Regulates Transcription of Msn2p-Dependent Stress-Responsive Genes

Acetyl-CoA Carboxylase Regulates Global Histone Acetylation

“…Denatured proteins were separated on a 15% denaturing polyacrylamide gel. Western blotting with antihistone H3 polyclonal antibody (ab1791; Abcam) at a dilution of 1:1000, anti-acetyl histone H3 (Lys-14) polyclonal antibody (acH3K14; 07-353, Upstate Biotechnology, Inc.) at a dilution of 1:500, and anti-hyperacetylated histone H4 polyclonal antibody (acH4K5,8,12,16; 06-946; Upstate Biotechnology, Inc.) at a dilution of 1:1000 was carried out as described previously (22). Western blots were quantified using NIH ImageJ software (ImageJ, National Institutes of Health, imagej.nih.gov).…”

“…Chromatin Immunoprecipitation and Quantitative Real Time PCR Analysis-In vivo chromatin cross-linking and immunoprecipitation were performed essentially as described (22,26,27). Immunoprecipitation was performed with the following antibodies: for anti-histone H3 (Abcam ab1791), antiacetyl-histone H3 (Lys-14) (Upstate Cell Signaling Solutions 07-353), and for anti-hyperacetylated histone H4 (Penta) (Upstate Cell Signaling Solutions 06-946).…”

Regulation of Inositol Metabolism Is Fine-tuned by Inositol Pyrophosphates in Saccharomyces cerevisiae*

“…Because of the difficulty of constructing a strain that can generate 5PP- IP 4 and IP 6 , but not IP 7 , it is difficult to elucidate the specific role of IP 7 in regulating INO1 transcription. Interestingly, deletion of PLC1, the gene encoding phospholipase C that hydrolyzes phosphatidylinositol 4,5-bisphosphate and generates IP 3 as precursors for inositol poly-/pyrophosphates, exhibited elevated INO1 expression (55,56). It is likely that regulation of INO1 gene expression and inositol biosynthesis is coordinated with phospholipase C activation in addition to the negative feedback circuit in response to exogenous inositol.…”

Yeast Phospholipase C Is Required for Normal Acetyl-CoA Homeostasis and Global Histone Acetylation

“…Following the general trend that promoters of highly transcribed genes are generally associated with increased histone acetylation (72)(73)(74), the acetylation of histones in ribosomal protein gene promoters by NuA4 is known to regulate their transcription (75,76). Our previous results showed that many ribosomal protein genes have decreased expression in plc1⌬ cells (44). Consistently with the notion that plc1⌬ cells display global histone hypoacetylation, the acetylation per nucleosome in the promoters of two ribosomal protein genes, RPL18B and RPS22B, was decreased in plc1⌬ cells in comparison with wild-type cells, and the decreased acetylation in plc1⌬ cells was suppressed by the hda1⌬ mutation (Fig.…”

“…Western Blotting-Denatured proteins were separated on 15% denaturing polyacrylamide gels and Western blotting with anti-histone H3 polyclonal antibody (ab1791; Abcam), antihistone H4 polyclonal antibody (2592; Cell Signaling), antiacetyl histone H3 (Lys-14) polyclonal antibody (acH3K14; 07-353, Upstate Biotechnology), anti-hyperacetylated histone H4 polyclonal antibody (acH4K5,8,12,16; 06-946; Upstate Biotechnology), anti-Htz1p polyclonal antibody (ab4626; Abcam), anti-acetyl-Htz1p (Lys-14) polyclonal antibody (07-719; Upstate), and anti-myc (A-14) polyclonal antibody (sc-789; Santa Cruz Biotechnology) was carried out as described previously (44). To confirm equivalent amounts of loaded proteins, the membranes were also probed5Ј with anti-Pgk1p monoclonal antibody 22C5 (A6457; Invitrogen).…”