There is growing consensus that deposition of insoluble deposits of betaA4 peptide cleaved from the amyloid precursor protein (APP) are central to the pathobiology of AD. Overproduction of APP and betaA4 can result from excess APP mRNA as in patients with Down's Syndrome (DS), APP homozygous transgenic animals and a subset of late onset AD patients. We have shown that APP mRNA abundance and downstream APP synthesis are controlled post-transcriptionally by at least two, 3' untranslated region regulatory elements. The 29 base element (29be) is 200 bases 3' from the stop codon and destabilizes APP mRNA. Recently, we discovered that APP mRNA contains a hitherto unknown stabilizing element located adjacent to and extending 50 bases 3' from the stop codon (50sc element or 50sce). This region is present uniquely in APP mRNAs. At least one cytoplasmic protein interacts specifically with the 50sce. Binding activity was detected in multiple cell lines and primary cells, suggesting widespread distribution. Deletion of the 50sc element accelerated APP mRNA degradation by up to 8 fold in transfected cells and reduced APP production by approximately the same amount. Endogenous, wild type APP mRNA and protein were dramatically reduced in SH-SY-5Y cells transfected with 50sce antisense oligonucleotides or decoy mRNAs containing a single copy of the sense 50sce. Thus we hypothesize that the 50sce also regulates the rate of APP mRNA decay and does so by specific interactions with cytoplasmic binding proteins. We also hypothesize that interference with the 50sc element will destabilize APP mRNA and reduce APP production. Therefore, we propose 1). To identify the essential nucleotides required for the stabilizing function of the 50sc element, 2). Determine if the 50sc element can function alone or does so in concert with other regions of the APP 3' UTR, 3). Identify, purify and clone the RNA binding proteins which recognize the 50sce, 4). Determine if blockade of the 50sc element destabilizes APP mRNA in transformed cells and primary neurons, 5). Produce a knock-in mouse lacking a functional 50sc element. In aggregate, these studies will help unravel how APP mRNA stability is regulated in cells and animal models. This knowledge may facilitate the development of anti-AP .

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG010675-14
Application #
6894290
Study Section
Special Emphasis Panel (ZRG1-MDCN-1 (01))
Program Officer
Snyder, Stephen D
Project Start
1991-09-29
Project End
2007-06-30
Budget Start
2005-07-01
Budget End
2006-06-30
Support Year
14
Fiscal Year
2005
Total Cost
$318,712
Indirect Cost
Name
University of Wisconsin Madison
Department
Pathology
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Westmark, Cara J; Malter, James S (2012) The regulation of AýýPP expression by RNA-binding proteins. Ageing Res Rev 11:450-9
Westmark, Cara J; Westmark, Pamela R; Malter, James S (2009) MPEP reduces seizure severity in Fmr-1 KO mice over expressing human Abeta. Int J Clin Exp Pathol 3:56-68
Broytman, Oleg; Westmark, Pamela R; Gurel, Zafer et al. (2009) Rck/p54 interacts with APP mRNA as part of a multi-protein complex and enhances APP mRNA and protein expression in neuronal cell lines. Neurobiol Aging 30:1962-74
Wang, Suqing; Simon, Brook P; Bennett, David A et al. (2007) The significance of Pin1 in the development of Alzheimer's disease. J Alzheimers Dis 11:13-23
Westmark, Cara J; Malter, James S (2007) FMRP mediates mGluR5-dependent translation of amyloid precursor protein. PLoS Biol 5:e52
Westmark, Pamela R; Shin, Hyun C; Westmark, Cara J et al. (2006) Decoy mRNAs reduce beta-amyloid precursor protein mRNA in neuronal cells. Neurobiol Aging 27:787-96
Westmark, Cara J; Gourronc, Francoise A; Bartleson, Virginia B et al. (2005) HuR mRNA ligands expressed after seizure. J Neuropathol Exp Neurol 64:1037-45
Broytman, Oleg; Malter, James S (2004) Anti-Abeta: The good, the bad, and the unforeseen. J Neurosci Res 75:301-6
Rapp, Peter R; Morrison, John H; Roberts, Jeffrey A (2003) Cyclic estrogen replacement improves cognitive function in aged ovariectomized rhesus monkeys. J Neurosci 23:5708-14
Malter, J S (2001) Regulation of mRNA stability in the nervous system and beyond. J Neurosci Res 66:311-6

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