Since our discovery of Arich element (ARE)-binding protein/degradation factor AUF1 in 1991, this protein family has been increasingly linked to many biological processes. These processes include cellular proliferation, inflammatory responses, transcription of cellular and viral genes, and maintenance of both telomeres and epigenetic integrity of the cell. Work from many laboratories, including the recent report of auf1-/- mice, has firmly established its participation in ARE-mediated mRNA degradation (AMD). During the prior funding period, we examined mechanisms of AUF1 action in AMD. Our key findings were that AUF1 acts within the context of a multi-protein complex that includes translation initiation factors and dual-functioning ARE-binding/heat shock proteins, and that AUF1 possesses an RNA chaperone-like activity that might control assembly of this protein complex. AUF1 thereby links the ARE-mRNP to the mRNA degradation machinery. We unexpectedly discovered that AUF1 controls translation in an ARE-dependent fashion, independently of its mRNA degradation function. Moreover, participation of AUF1 in translation versus mRNA degradation may be dictated by the specific ARE to which it is bound. Using MYC mRNA as a model, we found that AUF1 and other ARE-binding proteins work in opposition to set MYC translation rates. We also obtained evidence from mutagenesis studies that secondary structure within the MYC 3'UTR contributes to its translational control. As well, microRNA let-7g controls MYC expression and its annealing site is embedded within predicted regions of RNA secondary structure. Based upon these observations, our central hypothesis is that specific RNA-binding proteins and microRNAs, as well as 3'UTR secondary structure, conspire to set translation levels of MYC and other ARE-mRNA subsets. A corollary hypothesis is AREs may contain embedded regulatory codes dictating translation versus degradation (or both). We plan to utilize human cell culture systems and a combination of genetic and biochemical approaches to examine our hypotheses. Specifically, we will: (i) examine mechanisms of AUF1- directed translational activation;(ii) examine contributions of AUF1 to RNA structure, availability of the let-7g target site, and translation, and (iii) examine differential, AUF1-dependent association of trans-acting factors with degraded versus translationally-regulated ARE-mRNAs.

Public Health Relevance

The public health significance of these studies is highlighted by the fact that proper regulatory control of many ARE-mRNAs is essential, as their deregulation is central to formation of many tumors. Proper control of certain genes is important for normal cell division and development. When control of these critical genes is lost, cancer often results. We discovered a protein called AUF1, which is responsible for control of genes involved in cell division. We are studying the mechanisms by which AUF1 controls these genes, which will help us to understand processes by which normal cells can become cancer cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA052443-19
Application #
8264488
Study Section
Molecular Genetics C Study Section (MGC)
Program Officer
Strasburger, Jennifer
Project Start
1990-07-01
Project End
2013-06-30
Budget Start
2012-06-01
Budget End
2013-06-30
Support Year
19
Fiscal Year
2012
Total Cost
$324,449
Indirect Cost
$116,468
Name
University of Medicine & Dentistry of NJ
Department
Genetics
Type
Schools of Medicine
DUNS #
617022384
City
Piscataway
State
NJ
Country
United States
Zip Code
08854
Lin, Jing-Yi; Li, Mei-Ling; Brewer, Gary (2014) mRNA decay factor AUF1 binds the internal ribosomal entry site of enterovirus 71 and inhibits virus replication. PLoS One 9:e103827
Kishor, Aparna; Tandukar, Bishal; Ly, Yann V et al. (2013) Hsp70 is a novel posttranscriptional regulator of gene expression that binds and stabilizes selected mRNAs containing AU-rich elements. Mol Cell Biol 33:71-84
White, Elizabeth J F; Brewer, Gary; Wilson, Gerald M (2013) Post-transcriptional control of gene expression by AUF1: mechanisms, physiological targets, and regulation. Biochim Biophys Acta 1829:680-8
Wu, Xiangyue; Chesoni, Sandra; Rondeau, Gaelle et al. (2013) Combinatorial mRNA binding by AUF1 and Argonaute 2 controls decay of selected target mRNAs. Nucleic Acids Res 41:2644-58
Li, Mei-Ling; Defren, Jennifer; Brewer, Gary (2013) Hsp27 and F-box protein ýý-TrCP promote degradation of mRNA decay factor AUF1. Mol Cell Biol 33:2315-26
Xin, Hong; Brown, Julie A; Gong, Changning et al. (2012) Association of the von Hippel-Lindau protein with AUF1 and posttranscriptional regulation of VEGFA mRNA. Mol Cancer Res 10:108-20
Qi, Mei-Yan; Wang, Zhi-Zhang; Zhang, Zhuo et al. (2012) AU-rich-element-dependent translation repression requires the cooperation of tristetraprolin and RCK/P54. Mol Cell Biol 32:913-28
Wu, Xiangyue; Brewer, Gary (2012) The regulation of mRNA stability in mammalian cells: 2.0. Gene 500:10-21
Zhang, Zhuo; Qin, Yong-Wen; Brewer, Gary et al. (2012) MicroRNA degradation and turnover: regulating the regulators. Wiley Interdiscip Rev RNA 3:593-600
Liao, Baisong; Hu, Yan; Brewer, Gary (2011) RNA-binding protein insulin-like growth factor mRNA-binding protein 3 (IMP-3) promotes cell survival via insulin-like growth factor II signaling after ionizing radiation. J Biol Chem 286:31145-52

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