TY - JOUR
T1 - Ser 524 is a phosphorylation site in MUTYH and Ser 524 mutations alter 8-oxoguanine (OG)
T2 - A mismatch recognition
AU - Kundu, Sucharita
AU - Brinkmeyer, Megan K.
AU - Eigenheer, Richard A.
AU - David, Sheila S.
PY - 2010/10/5
Y1 - 2010/10/5
N2 - MUTYH-associated polyposis (MAP) is a colorectal cancer predisposition syndrome that is caused by inherited biallelic mutations in the base excision repair (BER) gene, MUTYH. MUTYH is a DNA glycosylase that removes adenine (A) misinserted opposite 8-oxo-7,8-dihydro-2′-deoxyguanosine (OG). In this work, wild type (WT) MUTYH overexpressed using a baculovirus-driven insect cell expression system (BEVS) provided significantly higher levels of enzyme compared to bacterial overexpression. The isolated MUTYH enzyme was analyzed for potential post-translational modifications using mass spectrometry. An in vivo phosphorylation site was validated at Serine 524, which is located in the C-terminal OG recognition domain within the proliferating cell nuclear antigen (PCNA) binding region. Characterization of the phosphomimetic (S524D) and phosphoablating (S524A) mutants together with the observation that Ser 524 can be phosphorylated suggest that this residue may play an important regulatory role in vivo by altering stability and OG:A mismatch affinity.
AB - MUTYH-associated polyposis (MAP) is a colorectal cancer predisposition syndrome that is caused by inherited biallelic mutations in the base excision repair (BER) gene, MUTYH. MUTYH is a DNA glycosylase that removes adenine (A) misinserted opposite 8-oxo-7,8-dihydro-2′-deoxyguanosine (OG). In this work, wild type (WT) MUTYH overexpressed using a baculovirus-driven insect cell expression system (BEVS) provided significantly higher levels of enzyme compared to bacterial overexpression. The isolated MUTYH enzyme was analyzed for potential post-translational modifications using mass spectrometry. An in vivo phosphorylation site was validated at Serine 524, which is located in the C-terminal OG recognition domain within the proliferating cell nuclear antigen (PCNA) binding region. Characterization of the phosphomimetic (S524D) and phosphoablating (S524A) mutants together with the observation that Ser 524 can be phosphorylated suggest that this residue may play an important regulatory role in vivo by altering stability and OG:A mismatch affinity.
KW - 8-Oxoguanine
KW - Adenine glycosylase
KW - MAP
KW - MUTYH
KW - Phosphorylation
KW - Post-translational modification
UR - http://www.scopus.com/inward/record.url?scp=77957304048&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77957304048&partnerID=8YFLogxK
U2 - 10.1016/j.dnarep.2010.07.002
DO - 10.1016/j.dnarep.2010.07.002
M3 - Article
C2 - 20724227
AN - SCOPUS:77957304048
VL - 9
SP - 1026
EP - 1037
JO - DNA Repair
JF - DNA Repair
SN - 1568-7864
IS - 10
ER -