New mortalin and histidyl tRNA synthetase isoforms point out a pitfall in proteomic analysis of Egr1 genetically modified mice.
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 pI of the molecular chaperone mortalin (mtHsp70/PBP74/Grp75) and the apparent disappearance of histidyl tRNA synthetase (HisRS). We found that the pI 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.
Chardonnet
Solenne
S
Institut de Biochimie et Biophysique Moléculaire et Cellulaire, UMR 8619, CNRS, Université Paris-Sud, Orsay Cedex, France. solenne.chardonnet@ibaic.u-psud.fr
Decottignies
Paulette
P
Amar
Laurence
L
Le Caer
Jean-Pierre
JP
Davis
Sabrina
S
Laroche
Serge
S
Le Maréchal
Pierre
P
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D051766:Early Growth Response Protein 1 / Q000235:genetics*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006643:Histidine-tRNA Ligase / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D020543:Proteome / Q000032:analysis*; D040901:Proteomics
D002352:Carrier Proteins; D051766:Early Growth Response Protein 1; C486109:Egr1 protein, mouse; D018840:HSP70 Heat-Shock Proteins; C083794:Hspa9 protein, mouse; D007527:Isoenzymes; D020543:Proteome; D006643:Histidine-tRNA Ligase