title: Structure and biochemical characterization of protein acetyltransferase from Sulfolobus solfataricus.
abstract: The Sulfolobus solfataricus protein acetyltransferase (PAT) acetylates ALBA, an abundant nonspecific DNA-binding protein, on Lys(16) to reduce its DNA affinity, and the Sir2 deacetylase reverses the modification to cause transcriptional repression. This represents a "primitive" model for chromatin regulation analogous to histone modification in eukaryotes. We report the 1.84-A crystal structure of PAT in complex with coenzyme A. The structure reveals homology to both prokaryotic GNAT acetyltransferases and eukaryotic histone acetyltransferases (HATs), with an additional "bent helix" proximal to the substrate binding site that might play an autoregulatory function. Investigation of active site mutants suggests that PAT does not use a single general base or acid residue for substrate deprotonation and product reprotonation, respectively, and that a diffusional step, such as substrate binding, may be rate-limiting. The catalytic efficiency of PAT toward ALBA is low relative to other acetyltransferases, suggesting that there may be better, unidentified substrates for PAT. The structural similarity of PAT to eukaryotic HATs combined with its conserved role in chromatin regulation suggests that PAT is evolutionarily related to the eukaryotic HATs.
authors.affiliation: Wistar Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Michael M
authors.initials: MM
authors.lastname: Brent
authors.forename: Ayaka
authors.initials: A
authors.lastname: Iwata
authors.forename: Juliana
authors.initials: J
authors.lastname: Carten
authors.forename: Kehao
authors.initials: K
authors.lastname: Zhao
authors.forename: Ronen
authors.initials: R
authors.lastname: Marmorstein
mesh_terms: D000107:Acetylation; D000123:Acetyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D003460:Crystallization; D004268:DNA-Binding Proteins / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D048229:Sulfolobus solfataricus / Q000201:enzymology* / Q000235:genetics
chemical_list: D019843:Archaeal Proteins; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D000123:Acetyltransferases