TY - JOUR
T1 - Mutagenesis of tGCN5 core region reveals two critical surface residues F90 and R140
AU - Mehta, Kinjal Rajesh
AU - Chan, Yan M.
AU - Lee, Man X.
AU - Yang, Ching Yao
AU - Voloshchuk, Natalya
AU - Montclare, Jin Kim
PY - 2010/9
Y1 - 2010/9
N2 - Tetrahymena General Control Non-Derepressor 5 (tGCN5) is a critical regulator of gene transcription via acetylation of histones. Since the acetylation ability has been attributed to the " core region" , we perform mutagenesis of residues within the tGCN5 " core region" in order to identify those critical for function and stability. Residues that do not participate in catalysis are identified, mutated and characterized for activity, structure and thermodynamic stability. Variants I107V, Q114L, A121T and A130S maintain the acetylation function relative to wild-type tGCN5, while variants F90Y, F112R and R140H completely abolish function. Of the three non-functional variants, since F112 is mutated into a non-homologous charged residue, a loss in function is expected. However, the remaining two variants are mutated into homologous residues, suggesting that F90 and R140 are critical for the activity of tGCN5. While mutation to homologous residue maintains acetylation of histone H3 for the majority of the variants, the two surface-exposed residues, F90 and R140, appear to be essential for tGCN5 function, structure or stability.
AB - Tetrahymena General Control Non-Derepressor 5 (tGCN5) is a critical regulator of gene transcription via acetylation of histones. Since the acetylation ability has been attributed to the " core region" , we perform mutagenesis of residues within the tGCN5 " core region" in order to identify those critical for function and stability. Residues that do not participate in catalysis are identified, mutated and characterized for activity, structure and thermodynamic stability. Variants I107V, Q114L, A121T and A130S maintain the acetylation function relative to wild-type tGCN5, while variants F90Y, F112R and R140H completely abolish function. Of the three non-functional variants, since F112 is mutated into a non-homologous charged residue, a loss in function is expected. However, the remaining two variants are mutated into homologous residues, suggesting that F90 and R140 are critical for the activity of tGCN5. While mutation to homologous residue maintains acetylation of histone H3 for the majority of the variants, the two surface-exposed residues, F90 and R140, appear to be essential for tGCN5 function, structure or stability.
KW - Histone
KW - Histone acetyltransferase (HAT)
KW - Homology
KW - Structural stability
KW - TGCN5
KW - Thermodynamic stability
UR - http://www.scopus.com/inward/record.url?scp=77956898817&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77956898817&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2010.08.069
DO - 10.1016/j.bbrc.2010.08.069
M3 - Article
C2 - 20732305
AN - SCOPUS:77956898817
SN - 0006-291X
VL - 400
SP - 363
EP - 368
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -