Cloning of Presenilin 2 cDNA and Construction of Vectors Carrying Effective Mutations in the Pathogenesis of Familial Alzheimer's Disease
Abstract
DOI:
10.26650/electrica.2018.12050
Alzheimer's
disease (AD) is a neurodegenerative disease and is identified by the detection
of amyloid-plaques and neurofibrillary tangles in the brain. Amyloid precursor
protein gene, presenilin 1 (PSEN1) gene, and presenilin 2 (PSEN2) gene are
responsible for this disease. PSEN2 and amyloid precursor protein (APP) gene
mutations are a much rarer cause of familial AD patients. This study aims to
clone the PSEN2 gene and create vectors with different mutations by directed
mutagenesis. As a result of the experiments, the PSEN2 cDNA was cloned between
the BamHI and KpnI cut-off points of the pBluescript II sk (+) vector. PSEN1
and PSEN2 homologs have a role in cell destiny decision and AD progress. We studied
some of the PSEN2 mutations (Ala252Thr and Pro334Arg) and provided expression
analysis in eukaryotic cell cultures. Amyloid β-protein (Aβ), which is produced
by endoproteolytic cleavage of the APP, is considered to play a role in AD.
While nominal concentration of Aβ40 is 10 times of Aβ42, the last peptide is
firmly linked to AD pathogenesis. Amyloid β-protein is generated by the
γ-secretase cleavage of APP onset and the progression of AD, and it is the
primary ingredient of the senile plaques. The Aβ42 dodecamer plays a central
role in AD. In future studies, it will be determined if there is an increase in
Aβ42 protein levels, and the effect on this early onset AD can be identified.
References
- Kumar A and Ekavali SA. A review on Alzheimer’s disease pathophysiology and its management: an update. Pharmacological Reports 2015; 67:195–203.
- Guo et al. Functional analyses of major cancer-related signaling pathways in Alzheimer's disease etiology. BBA - Reviews on Cancer 2017; 1868: 341–358.
- Plečkaitytė M. Alzheimer’s disease: a molecular mechanism, new hypotheses, and therapeutic strategies. Medicina (Kaunas) 2010; 46(1).
- Crews L and Masliah E. Molecular mechanisms of neurodegeneration in Alzheimer’s disease. Human Molecular Genetics 2010; 19(1).
- Yokes BM. Molecular genetics of Alzheimer's Disease. Journal of Cell and Molecular Biology 2007; 6(2): 73-97.
- St George-Hyslop HP and Petit A. Molecular biology and genetics of Alzheimer’s disease. C. R. Biologies 2004; 328: 119–130.
- Parvathy S and Buxbaum DJ. Molecular Genetics Of Alzheımer Disease. 2002; 83: 1199-1213.
- St George-Hyslop HP. Molecular Genetics of Alzheimer’s Disease. Biol Psychiatry. 2000;47:183–199.
- DeFina AP et al. Alzheimer’s Disease Clinical and Research Update for Health Care Practitioners. Journal of Aging Research. 2013.
- Korolev OI. Alzheimer’s Disease: A Clinical and Basic Science Review. Medical Student Research Journal. 2014; 04.
- Cai Y et al. Mutations in presenilin 2 and its implications in Alzheimer’s disease and other dementiaassociated disorders. Clinical Interventions in Aging. 2015:10 1163–1172.
- Schellenberg GD, Bird TD, Wijsman EM, et al. Genetic linkage evidence for a familial Alzheimer’s disease locus on chromosome 14. Science. 1992;258:668-671.
- Campion D, Flaman JM, Brice A, et al. Mutations of the presenilin I gene in families with early-onset Alzheimer’s disease. Hum Mol Genet. 1995;4:373- 377.
- Clark RF, Cruts M, Korenblat KM, et al. A yeast artificial chromosome contig from human chromosome 14q24 spanning the Alzheimer’s disease locus AD3. Hum Mol Genet. 1995;4:1347-1354.
- Kwok JB, Taddei K, Hallupp M, et al. Two novel (M233T and R278T) presenilin-1 mutations in early-onset Alzheimer’s disease pedigrees and preliminary evidence for association of presenilin-1 mutations with a novel phenotype. Neuroreport. 1997;8:1537-1542.
- Tysoe C, Galinsky D, Robinson D, et al. Apo E and Apo CI loci are associated with dementia in younger but not older late-onset cases. Dement Geriatr Cogn Disord. 1998;9:19119-19118.
- Beck JA, Mead S, Campbell TA, et al. Two-octapeptide repeat deletion of prion protein associated with rapidly progressive dementia. Neurology. 2001;57:354-356.
- Suzuki N, Cheung TT, Cai XD, et al. An increased percentage of long amyloid β protein secreted by familial amyloid β protein precursor (β APP717) mutants. Science. 1994;264:1336-1340.
- Taddei K, Fisher C, Laws SM, et al. Association between presenilin-1 Glu318Gly mutation and familial Alzheimer’s disease in the Australian population. Mol Psychiatry. 2002;7:776-781.
- Levy-Lahad E, Wasco W, Poorkaj P, et al. Candidate gene for the chromosome 1 familial Alzheimer’s disease locus. Science. 1995;269:973-977.
- Rogaev EI, Sherrington R, Rogaeva EA, et al. Familial Alzheimer's disease in kindreds with missense mutations in a gene on chromosome 1 related to the Alzheimer's disease type 3 gene. Nature. 1995; 376(6543):775-778.
- Stanga S et al. Specificity of presenilin-1- and presenilin-2-dependent γ-secretases towards substrate processing. J. Cell. Mol. Med. 2018; 22(2): 823-833.
- Ngo S and Guo Z. Key residues for the oligomerization of Aβ42 protein in Alzheimer’s disease. Biochemical and Biophysical Research Communications. 2011; 414: 512–516.
- Roychaudhuri R, Yang M, Hoshi MM, Teplow DB. Amyloid beta-protein assembly and Alzheimer disease. J Biol Chem. 2009; 284:4749-4753.
- Younkin SG. Evidence that AP42 is the real culprit in Alzheimer's disease. Ann Neurol. 1995; 37:287-288.
- Birnbaum HJ et al. Oxidative stress and altered mitochondrial protein expression in the absence of amyloid-β and tau pathology in iPSC-derived neurons from sporadic Alzheimer's disease patients. Stem Cell Research. 2018; 27:121-130.
- Zekanowski C et al. Mutations in presenilin 1, presenilin 2 and amyloid precursor protein genes in patients with early-onset Alzheimer’s disease in Poland. Experimental Neurology. 2003; 184: 991 – 996.
- Weggen S and Beher D. Molecular consequences of amyloid precursor protein and presenilin mutations causing autosomal-dominant Alzheimer’s disease. Alzheimer’s Research & Therapy. 2012; 4:9.
Cloning of Presenilin 2 cDNA and Construction of Vectors Carrying Effective Mutations in the Pathogenesis of Familial Alzheimer's Disease
Abstract
DOI: 10.26650/electrica.2018.12050
Alzheimer's disease (AD) is a neurodegenerative disease and is identified by the detection of amyloid-plaques and neurofibrillary tangles in the brain. Amyloid precursor protein gene, presenilin 1 (PSEN1) gene, and presenilin 2 (PSEN2) gene are responsible for this disease. PSEN2 and amyloid precursor protein (APP) gene mutations are a much rarer cause of familial AD patients. This study aims to clone the PSEN2 gene and create vectors with different mutations by directed mutagenesis. As a result of the experiments, the PSEN2 cDNA was cloned between the BamHI and KpnI cut-off points of the pBluescript II sk (+) vector. PSEN1 and PSEN2 homologs have a role in cell destiny decision and AD progress. We studied some of the PSEN2 mutations (Ala252Thr and Pro334Arg) and provided expression analysis in eukaryotic cell cultures. Amyloid β-protein (Aβ), which is produced by endoproteolytic cleavage of the APP, is considered to play a role in AD. While nominal concentration of Aβ40 is 10 times of Aβ42, the last peptide is firmly linked to AD pathogenesis. Amyloid β-protein is generated by the γ-secretase cleavage of APP onset and the progression of AD, and it is the primary ingredient of the senile plaques. The Aβ42 dodecamer plays a central role in AD. In future studies, it will be determined if there is an increase in Aβ42 protein levels, and the effect on this early onset AD can be identified.
References
- Kumar A and Ekavali SA. A review on Alzheimer’s disease pathophysiology and its management: an update. Pharmacological Reports 2015; 67:195–203.
- Guo et al. Functional analyses of major cancer-related signaling pathways in Alzheimer's disease etiology. BBA - Reviews on Cancer 2017; 1868: 341–358.
- Plečkaitytė M. Alzheimer’s disease: a molecular mechanism, new hypotheses, and therapeutic strategies. Medicina (Kaunas) 2010; 46(1).
- Crews L and Masliah E. Molecular mechanisms of neurodegeneration in Alzheimer’s disease. Human Molecular Genetics 2010; 19(1).
- Yokes BM. Molecular genetics of Alzheimer's Disease. Journal of Cell and Molecular Biology 2007; 6(2): 73-97.
- St George-Hyslop HP and Petit A. Molecular biology and genetics of Alzheimer’s disease. C. R. Biologies 2004; 328: 119–130.
- Parvathy S and Buxbaum DJ. Molecular Genetics Of Alzheımer Disease. 2002; 83: 1199-1213.
- St George-Hyslop HP. Molecular Genetics of Alzheimer’s Disease. Biol Psychiatry. 2000;47:183–199.
- DeFina AP et al. Alzheimer’s Disease Clinical and Research Update for Health Care Practitioners. Journal of Aging Research. 2013.
- Korolev OI. Alzheimer’s Disease: A Clinical and Basic Science Review. Medical Student Research Journal. 2014; 04.
- Cai Y et al. Mutations in presenilin 2 and its implications in Alzheimer’s disease and other dementiaassociated disorders. Clinical Interventions in Aging. 2015:10 1163–1172.
- Schellenberg GD, Bird TD, Wijsman EM, et al. Genetic linkage evidence for a familial Alzheimer’s disease locus on chromosome 14. Science. 1992;258:668-671.
- Campion D, Flaman JM, Brice A, et al. Mutations of the presenilin I gene in families with early-onset Alzheimer’s disease. Hum Mol Genet. 1995;4:373- 377.
- Clark RF, Cruts M, Korenblat KM, et al. A yeast artificial chromosome contig from human chromosome 14q24 spanning the Alzheimer’s disease locus AD3. Hum Mol Genet. 1995;4:1347-1354.
- Kwok JB, Taddei K, Hallupp M, et al. Two novel (M233T and R278T) presenilin-1 mutations in early-onset Alzheimer’s disease pedigrees and preliminary evidence for association of presenilin-1 mutations with a novel phenotype. Neuroreport. 1997;8:1537-1542.
- Tysoe C, Galinsky D, Robinson D, et al. Apo E and Apo CI loci are associated with dementia in younger but not older late-onset cases. Dement Geriatr Cogn Disord. 1998;9:19119-19118.
- Beck JA, Mead S, Campbell TA, et al. Two-octapeptide repeat deletion of prion protein associated with rapidly progressive dementia. Neurology. 2001;57:354-356.
- Suzuki N, Cheung TT, Cai XD, et al. An increased percentage of long amyloid β protein secreted by familial amyloid β protein precursor (β APP717) mutants. Science. 1994;264:1336-1340.
- Taddei K, Fisher C, Laws SM, et al. Association between presenilin-1 Glu318Gly mutation and familial Alzheimer’s disease in the Australian population. Mol Psychiatry. 2002;7:776-781.
- Levy-Lahad E, Wasco W, Poorkaj P, et al. Candidate gene for the chromosome 1 familial Alzheimer’s disease locus. Science. 1995;269:973-977.
- Rogaev EI, Sherrington R, Rogaeva EA, et al. Familial Alzheimer's disease in kindreds with missense mutations in a gene on chromosome 1 related to the Alzheimer's disease type 3 gene. Nature. 1995; 376(6543):775-778.
- Stanga S et al. Specificity of presenilin-1- and presenilin-2-dependent γ-secretases towards substrate processing. J. Cell. Mol. Med. 2018; 22(2): 823-833.
- Ngo S and Guo Z. Key residues for the oligomerization of Aβ42 protein in Alzheimer’s disease. Biochemical and Biophysical Research Communications. 2011; 414: 512–516.
- Roychaudhuri R, Yang M, Hoshi MM, Teplow DB. Amyloid beta-protein assembly and Alzheimer disease. J Biol Chem. 2009; 284:4749-4753.
- Younkin SG. Evidence that AP42 is the real culprit in Alzheimer's disease. Ann Neurol. 1995; 37:287-288.
- Birnbaum HJ et al. Oxidative stress and altered mitochondrial protein expression in the absence of amyloid-β and tau pathology in iPSC-derived neurons from sporadic Alzheimer's disease patients. Stem Cell Research. 2018; 27:121-130.
- Zekanowski C et al. Mutations in presenilin 1, presenilin 2 and amyloid precursor protein genes in patients with early-onset Alzheimer’s disease in Poland. Experimental Neurology. 2003; 184: 991 – 996.
- Weggen S and Beher D. Molecular consequences of amyloid precursor protein and presenilin mutations causing autosomal-dominant Alzheimer’s disease. Alzheimer’s Research & Therapy. 2012; 4:9.
Details
| Primary Language | English |
|---|---|
| Subjects | Engineering |
| Journal Section | Articles |
| Authors | |
| Publication Date | August 3, 2018 |
| Published in Issue | Year 2018 Volume: 18 Issue: 2 |
