welcome to oneFAPvoice
- a positively charged Familial Adenomatous Polyposis community.- join today!
- login
Functional Analysis of MUTYH Mutated Proteins Associated with Familial Adenomatous Polyposis
source: DNA repair
year: 2010
authors: D'Agostino V G, Minoprio A, Torreri P, Marinoni I, Bossa C, Petrucci T C, Albertini A M, Ranzani G N, Bignami M, Mazzei F
summary/abstract:The MUTYH DNA glycosylase specifically removes adenine misincorporated by replicative polymerases opposite the oxidized purine 8-oxo-7,8-dihydroguanine (8-oxoG). A defective protein activity results in the accumulation of G>T transversions because of unrepaired 8-oxoG:A mismatches. In humans, MUTYH germline mutations are associated with a recessive form of familial adenomatous polyposis and colorectal cancer predisposition (MUTYH-associated polyposis, MAP). Here we studied the repair capacity of the MUTYH variants R171W, E466del, 137insIW, Y165C and G382D, identified in MAP patients. Following expression and purification of human proteins from a bacterial system, we investigated MUTYH incision capacity on an 8-oxoG:A substrate by standard glycosylase assays. For the first time, we employed the surface plasmon resonance (SPR) technology for real-time recording of the association/dissociation of wild-type and MUTYH variants from an 8-oxoG:A DNA substrate. When compared to the wild-type protein, R171W, E466del and Y165C variants showed a severe reduction in the binding affinity towards the substrate, while 137insIW and G382D mutants manifested only a slight decrease mainly due to a slower rate of association. This reduced binding was always associated with impairment of glycosylase activity, with adenine removal being totally abrogated in R171W, E466del and Y165C and only partially reduced in 137insIW and G382D. Our findings demonstrate that SPR analysis is suitable to identify defective enzymatic behaviour even when mutant proteins display minor alterations in substrate recognition.
organization: University of PaviaDOI: 10.1016/j.dnarep.2010.03.008
read more full text source
expertly curated content related to this topic
-
Familial Adenomatous PolyposisFamilial adenomatous polyposis (FAP) is ...
-
Targeting Wnt Signaling in Colorectal Cancer. A Review in the Theme: Cell Signaling: Proteins, Pathways and Mechanis...The evolutionarily conserved Wnt signali...
-
Familial Adenomatous Polyposis of the ColonFamilial adenomatous polyposis (FAP) is ...
-
Colorectal Cancers Choosing SidesIn contrast to the majority of sporadic ...
-
An APC:WNT Counter-Current-Like Mechanism Regulates Cell Division along the Human Colonic Crypt Axis: A Mechanism th...APC normally down-regulates WNT signalin...
-
Global Quantitative Assessment of the Colorectal Polyp Burden in Familial Adenomatous Polyposis by Using a Web-based...BACKGROUND: Accurate measures of the to...
-
The Genetics of Familial Adenomatous Polyposis (FAP) and MutYH-Associated Polyposis (MAP)FAP is characterized by 100-1000s of ade...