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New mortalin and histidyl tRNA synthetase isoforms point out a pitfall in proteomic analysis of Egr1 genetically modified mice
Author(s) -
Chardonnet Solenne,
Decottignies Paulette,
Amar Laurence,
Le Caer JeanPierre,
Davis Sabrina,
Laroche Serge,
Le Maréchal Pierre
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200600513
Subject(s) - egr1 , gene isoform , biology , gene , complementary dna , genetics , microbiology and biotechnology , haplotype , allele , exon , gene expression
Egr1 ( Zif268 ) is an immediate early gene encoding an inducible transcription factor involved in synaptic plasticity and several forms of memory in rodents. Using 2‐DE and MS, we compared proteomes of hippocampal subregions and cortex in Egr1‐deficient and wild‐type littermates. Two significant differences were identified: a shift in the p I of the molecular chaperone mortalin (mtHsp70/PBP74/Grp75) and the apparent disappearance of histidyl tRNA synthetase (HisRS). We found that the p I shift for mortalin in Egr1‐deficient mice was caused by a difference in protein sequence: D626G. Using cDNA sequencing, we demonstrated for both mortalin and HisRS that protein differences were not due to a lack of Egr1 but to DNA polymorphism between the C57Bl/6J and 129/Sv strains used to generate the Egr1‐deficient mice. Our results show that mortalin and HisRS genes, which map closely to the Egr1 locus, have conserved the 129/Sv haplotype despite numerous back‐crossing of the null mice progeny with C57Bl/6J animals. This demonstrates that allelic differences between mouse strains can introduce variations in differential proteomic analyses of genetically modified organisms. Finally, we report the identification of new isoforms of HisRS and mortalin (mot‐3) encoded by the 129/Sv haplotype.