Linked Life Data

11461190

Source:http://linkedlifedata.com/resource/pubmed/id/11461190

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
2001-7-19
pubmed:abstractText
Phenylketonuria (PKU) is caused by mutations in the phenylalanine hydroxylase gene (PAH), while mutations in genes encoding the two enzymes (dihydropteridine reductase, DHPR, and pterin-4-alpha-carbinolamine dehydratase, PCD) required for recycling of its cofactor, tetrahydrobiopterin (BH(4)), cause other rarer disease forms of hyperphenylalaninemia. We have applied a yeast two-hybrid method, in which protein--protein interactions are measured by four reporter gene constructs, to the analysis of six PKU-associated PAH missense mutations (F39L, K42I, L48S, I65T, A104D, and R157N). By studying homomeric interactions between mutant PAH subunits, we show that this system is capable of detecting quite subtle aberrations in PAH oligomerization caused by missense mutations and that the observed results generally correlate with the severity of the mutation as determined by other expression systems. The mutant PAH subunits are also shown in this system to be able to interact with wild-type PAH subunits, pointing to an explanation for apparent dominant negative effects previously observed in obligate heterozygotes for PKU mutations. Based on our findings, the applications and limitations of two-hybrid approaches in understanding mechanisms by which PAH missense mutations exert their pathogenic effects are discussed. We have also used this technique to demonstrate homomeric interactions between wild-type DHPR subunits and between wild-type PCD subunits. These data provide a basis for functional studies on HPA-associated mutations affecting these enzymes.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
1096-7192
pubmed:author
pubmed:copyrightInfo
Copyright 2001 Academic Press.
pubmed:issnType
Print
pubmed:volume
73
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
230-8
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:11461190-Alleles, pubmed-meshheading:11461190-Biopterin, pubmed-meshheading:11461190-DNA, Complementary, pubmed-meshheading:11461190-Dihydropteridine Reductase, pubmed-meshheading:11461190-Dimerization, pubmed-meshheading:11461190-Genes, Reporter, pubmed-meshheading:11461190-Heterozygote, pubmed-meshheading:11461190-Humans, pubmed-meshheading:11461190-Hydro-Lyases, pubmed-meshheading:11461190-Mutation, pubmed-meshheading:11461190-Phenylalanine, pubmed-meshheading:11461190-Phenylalanine Hydroxylase, pubmed-meshheading:11461190-Plasmids, pubmed-meshheading:11461190-Protein Binding, pubmed-meshheading:11461190-Protein Biosynthesis, pubmed-meshheading:11461190-Protein Structure, Tertiary, pubmed-meshheading:11461190-Transcription, Genetic, pubmed-meshheading:11461190-Two-Hybrid System Techniques
pubmed:year
2001
pubmed:articleTitle
Homomeric and heteromeric interactions between wild-type and mutant phenylalanine hydroxylase subunits: evaluation of two-hybrid approaches for functional analysis of mutations causing hyperphenylalaninemia.
pubmed:affiliation
deBelle Laboratory for Biochemical Genetics, Montreal Children's Hospital, Quebec, Canada. paula.walters@muhc.mcgill.ca
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't