Transcriptional activation of heat shock factor HSF1 probed by phosphopeptide analysis of factor 32P-labeled in vivo.

The Journal of Biological Chemistry
W XiaR Voellmy

Abstract

Mapping of tryptic phosphopeptides of heat shock factor 1 (HSF1) from non-stressed or moderately heat-stressed HeLa cells, labeled in vivo by [32P]orthophosphate, revealed four major phosphopeptides A to D. Heat stress drastically increased phosphopeptide signals. To identify target peptides and amino acids and to correlate phosphorylation and transactivation function, phosphopeptide maps were produced of LexA-human HSF1 chimeras and mutant derivatives thereof, and transactivation activities of original and mutant chimeras were compared. LexA-HSF1 chimeras were previously shown to be regulated identically to HSF1, except that they transactivate promoters with LexA-binding sites instead of hsp promoters. The patterns of phosphopeptides of LexA-HSF1 and endogenous HSF1 were similar. Analysis of single residue substitutions suggested that phosphopeptide C is peptide VKEEPPSPPQSPR (297-309) phosphorylated on Ser-307 but not Ser-303. Substitution of Ser-307 but not Ser-303 caused deregulation of factor activity. Mapping of several constitutively active chimeras associated unphosphorylated peptide C with the transcriptionally active HSF1 conformation, suggesting that dephosphorylation of this peptide (at Ser-307) may either be an int...Continue Reading

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Citations

Aug 31, 2001·Progress in Neurobiology·C A Grimes, R S Jope
Dec 7, 2002·Trends in Biochemical Sciences·Carina I HolmbergLea Sistonen
Jul 16, 2010·Nature Reviews. Molecular Cell Biology·Malin AkerfeltLea Sistonen
Mar 12, 2005·BMC Biochemistry·Toumy GuettoucheRichard Voellmy
Mar 23, 2011·Annual Review of Biochemistry·Julius Anckar, Lea Sistonen
Sep 1, 2004·Biochemical and Biophysical Research Communications·Alex MammenCheil Moon
Jun 24, 2004·Progress in Nucleic Acid Research and Molecular Biology·Richard Voellmy
Sep 28, 2001·The Journal of Biological Chemistry·K F WinklhoferJ Tatzelt
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Apr 14, 2021·GeroScience·Rachana TrivediDonald A Jurivich
Mar 21, 2003·Biochemical and Biophysical Research Communications·Roland S HilgarthKevin D Sarge

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