Molecular basis of competition between HSF2 and catalytic subunit for binding to the PR65/A subunit of PP2A.
We recently identified the existence of a novel interaction between heat shock transcription factor 2 (HSF2) and the PR65/A subunit of protein phosphatase 2A (PP2A) and showed that HSF2 is able to compete with the PP2A catalytic subunit for binding to PR65. To elucidate the mechanistic basis of this competition between HSF2 and catalytic subunit at the molecular level we have sought to characterize sequences within PR65 that are important for interaction with HSF2. The results identify the intra-repeat loop within HEAT repeat 11 of PR65 as critical for interaction with HSF2. Analysis of point mutants within this loop region of PR65 identify lysine 416 as a residue critical for interaction with HSF2. Interestingly, this same lysine residue of PR65 is important for its binding to catalytic subunit. These results suggest that HSF2's ability to interfere with catalytic subunit binding to PR65 is due to competition between HSF2 and catalytic subunit for at least one amino acid residue of PR65, lysine 416. These data support the hypothesis that HSF2 represents a new type of PP2A regulatory protein.
Hong
Y
Y
Department of Biochemistry, Chandler Medical Center, University of Kentucky, Lexington 40536, USA.
Lubert
E J
EJ
Rodgers
D W
DW
Sarge
K D
KD
D000595:Amino Acid Sequence; D001483:Base Sequence; D001667:Binding, Competitive; D020134:Catalytic Domain / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010749:Phosphoprotein Phosphatases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017354:Point Mutation; D054648:Protein Phosphatase 2; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D017931:DNA Primers; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors; C070389:HSF2 protein, human; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D008239:Lysine