The function and fate of any given cell is determined by the gene expression profile of that cell. While transcription plays a key role in determining gene expression, post-transcriptional regulatory events also are of vital importance in determining the spatial and temporal pattern of gene expression. One crucial post- transcriptional step is addition of a polyadenosine or poly(A) tail following 3'-end cleavage of mRNA. This poly(A) tail modulates numerous events including mRNA export from the nucleus, translation, transport and turnover, largely via the recruitment of polyadenosine RNA binding proteins (Pabs). Polyadenylation of additional classes of RNA has also been implicated in quality control. The critical importance of proper polyadenylation and Pab protein function is evident in the number of human diseases that arise due to mutations in genes encoding these proteins. In the previous funding cycle, we collaborated to identify mutations in the human ZC3H14 gene, which encodes a ubiquitously expressed, evolutionarily conserved, zinc finger, nuclear polyadenosine RNA binding protein (Pab), that cause an autosomal recessive form of non- syndromic intellectual disability (previously termed mental retardation). These patients have severely impaired brain function with IQs in the range of 35-50 as compared to an average adult IQ range of 90-110. Although little is known about the function of ZC3H14, studies of the budding yeast (Nab2) and Drosophila counterparts (dNab2) provide compelling evidence that this class of proteins plays an evolutionarily conserved role in regulating poly(A) tail length. The broad, long-term objective of this proposal is to define how poly(A) tail length is regulated by the zinc finger Pabs to understand both the critical role in post-transcriptional regulation of gene expression and the mechanisms underlying neuronal defects in affected patients. This proposal exploits exciting preliminary data collected through studies of this evolutionarily conserved class of proteins in budding yeast, Drosophila, and cultured neuronal cells capitalizing on established tools and models. These studies strongly implicate the complex of 3'-5 riboexonucleases known as the nuclear exosome in cooperating with Nab2 to regulate poly(A) tail length. Based on preliminary data, we will test the hypothesis that the Nab2 class of RNA binding proteins cooperates with the exosome to regulate poly(A) tail length of RNA thus ensuring proper neuronal function. This hypothesis will be tested through three complementary Specific Aims that seek to: 1) define a molecular mechanism for Nab2/ZC3H14 in regulating poly(A) tail length (Aim 1);extend these studies into our established Drosophila model to link molecular mechanisms to neuronal function (Aim 2);and finally determine whether specific RNAs or classes of RNAs accumulate extended poly(A) tails upon loss of Nab2/ZC3H14 function (Aim 3). Successful completion of these aims will provide insight into a critical point in post-transcriptional regulation of gene expression mediated by a recently described family of Pab proteins and also lend insight into the molecular defects underlying intellectual disability in patients.
This proposal seeks to understand the function of a protein that we recently found is altered in patients who have a disease called intellectual disability where their brain function is severely impaired (a disease previously termed mental retardation). The type of protein that is defective in these patients binds to the end or poly(A) tail of the messenger sequences or mRNAs that carry information from the genetic material in the cell nucleus to the cytoplasm where the information can be decoded. These proteins are very important for regulating how that information is read and thus understanding their function will provide important information both about how cells decode genes and about what is wrong in the brains of patients who suffer from this form of intellectual disability.
|Wigington, Callie P; Williams, Kathryn R; Meers, Michael P et al. (2014) Poly(A) RNA-binding proteins and polyadenosine RNA: new members and novel functions. Wiley Interdiscip Rev RNA 5:601-22|
|Kelly, Seth M; Leung, Sara W; Pak, ChangHui et al. (2014) A conserved role for the zinc finger polyadenosine RNA binding protein, ZC3H14, in control of poly(A) tail length. RNA 20:681-8|
|Pak, Changhui; Garshasbi, Masoud; Kahrizi, Kimia et al. (2011) Mutation of the conserved polyadenosine RNA binding protein, ZC3H14/dNab2, impairs neural function in Drosophila and humans. Proc Natl Acad Sci U S A 108:12390-5|
|Kerr, Shana C; Azzouz, Nowel; Fuchs, Stephen M et al. (2011) The Ccr4-Not complex interacts with the mRNA export machinery. PLoS One 6:e18302|
|Swartzlander, Dan B; Griffiths, Lyra M; Lee, Joan et al. (2010) Regulation of base excision repair: Ntg1 nuclear and mitochondrial dynamic localization in response to genotoxic stress. Nucleic Acids Res 38:3963-74|
|Carmody, Sean R; Tran, Elizabeth J; Apponi, Luciano H et al. (2010) The mitogen-activated protein kinase Slt2 regulates nuclear retention of non-heat shock mRNAs during heat shock-induced stress. Mol Cell Biol 30:5168-79|
|Kerr, Shana C; Corbett, Anita H (2010) Should INO stay or should INO Go: a DNA "zip code" mediates gene retention at the nuclear pore. Mol Cell 40:3-5|
|Leung, Sara W; Apponi, Luciano H; Cornejo, Omar E et al. (2009) Splice variants of the human ZC3H14 gene generate multiple isoforms of a zinc finger polyadenosine RNA binding protein. Gene 439:71-8|
|Griffiths, Lyra M; Swartzlander, Dan; Meadows, Kellen L et al. (2009) Dynamic compartmentalization of base excision repair proteins in response to nuclear and mitochondrial oxidative stress. Mol Cell Biol 29:794-807|
|Kelly, Seth M; Corbett, Anita H (2009) Messenger RNA export from the nucleus: a series of molecular wardrobe changes. Traffic 10:1199-208|
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