当前位置： SCI文献检索 > Computational and Structural Biotechnology Journal期刊下所有文献 > Characterization of ENU-induced Mutations in Red Blood Cell Structural Proteins.

Characterization of ENU-induced Mutations in Red Blood Cell Structural Proteins.

Abstract：

:Murine models with modified gene function as a result of N-ethyl-N-nitrosourea (ENU) mutagenesis have been used to study phenotypes resulting from genetic change. This study investigated genetic factors associated with red blood cell (RBC) physiology and structural integrity that may impact on blood component storage and transfusion outcome. Forward and reverse genetic approaches were employed with pedigrees of ENU-treated mice using a homozygous recessive breeding strategy. In a "forward genetic" approach, pedigree selection was based upon identification of an altered phenotype followed by exome sequencing to identify a causative mutation. In a second strategy, a "reverse genetic" approach based on selection of pedigrees with mutations in genes of interest was utilised and, following breeding to homozygosity, phenotype assessed. Thirty-three pedigrees were screened by the forward genetic approach. One pedigree demonstrated reticulocytosis, microcytic anaemia and thrombocytosis. Exome sequencing revealed a novel single nucleotide variation (SNV) in Ank1 encoding the RBC structural protein ankyrin-1 and the pedigree was designated Ank1(EX34). The reticulocytosis and microcytic anaemia observed in the Ank1(EX34) pedigree were similar to clinical features of hereditary spherocytosis in humans. For the reverse genetic approach three pedigrees with different point mutations in Spnb1 encoding RBC protein spectrin-1β, and one pedigree with a mutation in Epb4.1, encoding band 4.1 were selected for study. When bred to homozygosity two of the spectrin-1β pedigrees (a, b) demonstrated increased RBC count, haemoglobin (Hb) and haematocrit (HCT). The third Spnb1 mutation (spectrin-1β c) and mutation in Epb4.1 (band 4.1) did not significantly affect the haematological phenotype, despite these two mutations having a PolyPhen score predicting the mutation may be damaging. Exome sequencing allows rapid identification of causative mutations and development of databases of mutations predicted to be disruptive. These tools require further refinement but provide new approaches to the study of genetically defined changes that may impact on blood component storage and transfusion outcome.

journal_name

Comput Struct Biotechnol J

journal_title

Computational and structural biotechnology journal

authors

Kildey K,Flower RL,Tran TV,Tunningley R,Harris J,Dean MM

doi

10.5936/csbj.201303012

subject

Has Abstract

pub_date

2013-09-23 00:00:00

pages

e201303012

issn

2001-0370

pii

CSBJ-6-e201303012

journal_volume

pub_type

杂志文章

在线工具

Characterization of ENU-induced Mutations in Red Blood Cell Structural Proteins.