Experimental and Computational Considerations in the Study of RNA-Binding Protein-RNA Interactions.

After an RNA is transcribed, it undergoes a variety of processing steps that can change the encoded protein sequence (through alternative splicing and RNA editing), regulate the stability of the RNA, and control subcellular localization, timing, and rate of translation. The recent explosion in genomics techniques has enabled transcriptome-wide profiling of RNA processing in an unbiased manner. However, it has also brought with it both experimental challenges in developing improved methods to probe distinct processing steps, as well as computational challenges in data storage, processing, and analysis tools to enable large-scale interpretation in the genomics era. In this chapter we review experimental techniques and challenges in profiling various aspects of RNA processing, as well as recent efforts to develop analyses integrating multiple data sources and techniques to infer RNA regulatory networks.

[1]  R. Darnell,et al.  Mapping in vivo protein-RNA interactions at single-nucleotide resolution from HITS-CLIP data , 2011, Nature Biotechnology.

[2]  Lisa A. McPherson,et al.  AP2alpha and AP2gamma: a comparison of binding site specificity and trans-activation of the estrogen receptor promoter and single site promoter constructs. , 1999, Nucleic acids research.

[3]  Martin J. Simard,et al.  Argonaute proteins: key players in RNA silencing , 2008, Nature Reviews Molecular Cell Biology.

[4]  K. Neugebauer,et al.  How cells get the message: dynamic assembly and function of mRNA–protein complexes , 2013, Nature Reviews Genetics.

[5]  Phillip A Sharp,et al.  Predictive Identification of Exonic Splicing Enhancers in Human Genes , 2002, Science.

[6]  Ron Shamir,et al.  Accurate identification of alternatively spliced exons using support vector machine , 2005, Bioinform..

[7]  Cole Trapnell,et al.  Ultrafast and memory-efficient alignment of short DNA sequences to the human genome , 2009, Genome Biology.

[8]  David G Hendrickson,et al.  Differential analysis of gene regulation at transcript resolution with RNA-seq , 2012, Nature Biotechnology.

[9]  Stephanie C Huelga,et al.  Divergent roles of ALS-linked proteins FUS/TLS and TDP-43 intersect in processing long pre-mRNAs , 2012, Nature Neuroscience.

[10]  Brendan J. Frey,et al.  Deciphering the splicing code , 2010, Nature.

[11]  Thomas R. Gingeras,et al.  STAR: ultrafast universal RNA-seq aligner , 2013, Bioinform..

[12]  Nicholas T. Ingolia,et al.  Genome-Wide Analysis in Vivo of Translation with Nucleotide Resolution Using Ribosome Profiling , 2009, Science.

[13]  Uwe Ohler,et al.  FMR1 targets distinct mRNA sequence elements to regulate protein expression , 2012, Nature.

[14]  Robert B Darnell,et al.  Nova-1 Regulates Neuron-Specific Alternative Splicing and Is Essential for Neuronal Viability , 2000, Neuron.

[15]  D. Black,et al.  Developmental Control of CaV1.2 L-Type Calcium Channel Splicing by Fox Proteins , 2009, Molecular and Cellular Biology.

[16]  R. Darnell,et al.  The neuronal RNA-binding protein Nova-2 is implicated as the autoantigen targeted in POMA patients with dementia. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Jonathan S. Weissman,et al.  Principles of ER cotranslational translocation revealed by proximity-specific ribosome profiling , 2014, Science.

[18]  Tomaso Poggio,et al.  Identification and analysis of alternative splicing events conserved in human and mouse. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Gene W. Yeo,et al.  Genome-wide analysis reveals SR protein cooperation and competition in regulated splicing. , 2013, Molecular cell.

[20]  Helga Thorvaldsdóttir,et al.  Integrative Genomics Viewer , 2011, Nature Biotechnology.

[21]  Shifeng Xue,et al.  Ribosome-Mediated Specificity in Hox mRNA Translation and Vertebrate Tissue Patterning , 2011, Cell.

[22]  Julia Salzman,et al.  Proteome-Wide Search Reveals Unexpected RNA-Binding Proteins in Saccharomyces cerevisiae , 2010, PloS one.

[23]  Jernej Ule,et al.  Understanding splicing regulation through RNA splicing maps , 2011, Trends in genetics : TIG.

[24]  J. Ule,et al.  iCLIP reveals the function of hnRNP particles in splicing at individual nucleotide resolution , 2010, Nature Structural &Molecular Biology.

[25]  S. Batzoglou,et al.  Genome-Wide Analysis of Transcription Factor Binding Sites Based on ChIP-Seq Data , 2008, Nature Methods.

[26]  Hua Shi,et al.  RNA aptamers that functionally interact with green fluorescent protein and its derivatives , 2011, Nucleic acids research.

[27]  Gene W. Yeo,et al.  An RNA code for the FOX2 splicing regulator revealed by mapping RNA-protein interactions in stem cells , 2009, Nature Structural &Molecular Biology.

[28]  Eric T. Wang,et al.  Transcriptome-wide Regulation of Pre-mRNA Splicing and mRNA Localization by Muscleblind Proteins , 2012, Cell.

[29]  T. Babak,et al.  A quantitative atlas of polyadenylation in five mammals , 2012, Genome research.

[30]  Jeroen Krijgsveld,et al.  The RNA-binding protein repertoire of embryonic stem cells , 2013, Nature Structural &Molecular Biology.

[31]  C. Burge,et al.  Global analyses of UPF1 binding and function reveal expanded scope of nonsense-mediated mRNA decay , 2013, Genome research.

[32]  Jernej Ule,et al.  CLIP Identifies Nova-Regulated RNA Networks in the Brain , 2003, Science.

[33]  Gene W. Yeo,et al.  Robust transcriptome-wide discovery of RNA binding protein binding sites with enhanced CLIP (eCLIP) , 2016, Nature Methods.

[34]  S. Jaffrey,et al.  RNA Mimics of Green Fluorescent Protein , 2011, Science.

[35]  Tamer Kahveci,et al.  Accessed Terms of Use , 2022 .

[36]  M. Selbach,et al.  Global quantification of mammalian gene expression control , 2011, Nature.

[37]  Ron Shamir,et al.  A non-EST-based method for exon-skipping prediction. , 2004, Genome research.

[38]  Nicholas T. Ingolia,et al.  Ribosome Profiling of Mouse Embryonic Stem Cells Reveals the Complexity and Dynamics of Mammalian Proteomes , 2011, Cell.

[39]  J. Cate,et al.  eIF3 targets cell proliferation mRNAs for translational activation or repression , 2015, Nature.

[40]  Scott B. Dewell,et al.  Transcriptome-wide Identification of RNA-Binding Protein and MicroRNA Target Sites by PAR-CLIP , 2010, Cell.

[41]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[42]  Tyson A. Clark,et al.  Aberrant alternative splicing and extracellular matrix gene expression in mouse models of myotonic dystrophy , 2010, Nature Structural &Molecular Biology.

[43]  J. Castle,et al.  Genome-Wide Survey of Human Alternative Pre-mRNA Splicing with Exon Junction Microarrays , 2003, Science.

[44]  B. Frey,et al.  The human splicing code reveals new insights into the genetic determinants of disease , 2015, Science.

[45]  Peter J. Shepard,et al.  Complex and dynamic landscape of RNA polyadenylation revealed by PAS-Seq. , 2011, RNA.

[46]  Karl-Heinz Glatting,et al.  Genome-wide prediction of splice-modifying SNPs in human genes using a new analysis pipeline called AASsites , 2011, BMC Bioinformatics.

[47]  A. Wakamatsu,et al.  Genome-wide determination of RNA stability reveals hundreds of short-lived noncoding transcripts in mammals , 2012, Genome research.

[48]  Daniel Herschlag,et al.  Diverse RNA-Binding Proteins Interact with Functionally Related Sets of RNAs, Suggesting an Extensive Regulatory System , 2008, PLoS biology.

[49]  C. Smith,et al.  Short hairpin RNA-mediated gene silencing. , 2013, Methods in molecular biology.

[50]  R. Darnell,et al.  The neuronal RNA binding protein Nova-1 recognizes specific RNA targets in vitro and in vivo , 1997, Molecular and cellular biology.

[51]  R. Schneider,et al.  Nuclear Import and Export Functions in the Different Isoforms of the AUF1/Heterogeneous Nuclear Ribonucleoprotein Protein Family* , 2003, Journal of Biological Chemistry.

[52]  E. Wang,et al.  Analysis and design of RNA sequencing experiments for identifying isoform regulation , 2010, Nature Methods.

[53]  Sarath Chandra Janga,et al.  A Screen for RNA-Binding Proteins in Yeast Indicates Dual Functions for Many Enzymes , 2010, PloS one.

[54]  Shihao Shen,et al.  MADS+: discovery of differential splicing events from Affymetrix exon junction array data , 2009, Bioinform..

[55]  Liang Chen,et al.  Statistical and Computational Studies on Alternative Splicing , 2011, Handbook of Statistical Bioinformatics.

[56]  Gene W. Yeo,et al.  Discovery and Analysis of Evolutionarily Conserved Intronic Splicing Regulatory Elements , 2007, PLoS Genetics.

[57]  T. Cooper,et al.  Muscleblind proteins regulate alternative splicing , 2004, The EMBO journal.

[58]  D. Cleveland,et al.  TDP-43 and FUS/TLS: emerging roles in RNA processing and neurodegeneration. , 2010, Human molecular genetics.

[59]  T. Borodina,et al.  Transcriptome analysis by strand-specific sequencing of complementary DNA , 2009, Nucleic acids research.

[60]  D. Bunka,et al.  An RNA Aptamer Provides a Novel Approach for the Induction of Apoptosis by Targeting the HPV16 E7 Oncoprotein , 2013, PloS one.

[61]  David Z. Chen,et al.  Architecture of the human regulatory network derived from ENCODE data , 2012, Nature.

[62]  Chris Williams,et al.  RNA-SeQC: RNA-seq metrics for quality control and process optimization , 2012, Bioinform..

[63]  R. Darnell,et al.  Sequence-Specific RNA Binding by a Nova KH Domain Implications for Paraneoplastic Disease and the Fragile X Syndrome , 2000, Cell.

[64]  M. Cleary,et al.  Comparative assessment of siRNA and shRNA off target effects: what is slowing clinical development , 2009, Cancer Gene Therapy.

[65]  Eric T. Wang,et al.  Alternative Isoform Regulation in Human Tissue Transcriptomes , 2008, Nature.

[66]  Norman E. Davey,et al.  Insights into RNA Biology from an Atlas of Mammalian mRNA-Binding Proteins , 2012, Cell.

[67]  Eric T. Wang,et al.  MBNL proteins repress ES-cell-specific alternative splicing and reprogramming , 2013, Nature.

[68]  A. Mortazavi,et al.  Genome-Wide Mapping of in Vivo Protein-DNA Interactions , 2007, Science.

[69]  F. Pontén,et al.  Correlations between RNA and protein expression profiles in 23 human cell lines , 2009, BMC Genomics.

[70]  Raymond K. Auerbach,et al.  Integrative Analysis of the Caenorhabditis elegans Genome by the modENCODE Project , 2010, Science.

[71]  Brendan J. Frey,et al.  Inferring global levels of alternative splicing isoforms using a generative model of microarray data , 2006, Bioinform..

[72]  Gos Micklem,et al.  Supporting Online Material Materials and Methods Figs. S1 to S50 Tables S1 to S18 References Identification of Functional Elements and Regulatory Circuits by Drosophila Modencode , 2022 .

[73]  Istvan Mody,et al.  The splicing regulator Rbfox1 (A2BP1) controls neuronal excitation in the mammalian brain , 2011, Nature Genetics.

[74]  Gene W. Yeo,et al.  Target Discrimination in Nonsense-Mediated mRNA Decay Requires Upf1 ATPase Activity. , 2015, Molecular cell.

[75]  M. Gorospe,et al.  Concurrent versus individual binding of HuR and AUF1 to common labile target mRNAs , 2004, The EMBO journal.

[76]  Gene W. Yeo,et al.  Integrative genome‐wide analysis reveals cooperative regulation of alternative splicing by hnRNP proteins , 2012, Cell reports.

[77]  Steven E Brenner,et al.  Genome-wide Analysis of Alternative Pre-mrna Splicing and Rna-binding Specificities of the Drosophila Hnrnp A/b Family Members , 2022 .

[78]  B. Blencowe,et al.  An RNA map predicting Nova-dependent splicing regulation , 2006, Nature.

[79]  Tom H. Pringle,et al.  The human genome browser at UCSC. , 2002, Genome research.

[80]  D. Tollervey,et al.  The Many Pathways of RNA Degradation , 2009, Cell.

[81]  Tyson A. Clark,et al.  HITS-CLIP yields genome-wide insights into brain alternative RNA processing , 2008, Nature.

[82]  T. Cooper Use of minigene systems to dissect alternative splicing elements. , 2005, Methods.