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Re-analysis of: ‘CSF proteome mapping: Human cerebrospinal fluid LC-MS/MS, Part 2’ (PXD000652) to demonstrate the ProteoAnnotator software for mapping identifications to Ensembl genomes

ABSTRACT: We have performed a re-analysis of PXD000652 - a human proteome CSF fluid sample, originally queried versus UniProt. We have used our ProteoAnnotator package to re-search the data against Ensembl Human build 76 to demonstrate the capability of the pipeline to perform genome mapping on a large scale.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Dr Andy Jones

PROVIDER: MSV000080193 | MassIVE |

SECONDARY ACCESSION(S): PXD001390

REPOSITORIES: MassIVE

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ProteoAnnotator--open source proteogenomics annotation software supporting PSI standards.

Ghali Fawaz F Krishna Ritesh R Perkins Simon S Collins Andrew A Xia Dong D Wastling Jonathan J Jones Andrew R AR

Proteomics 20141119 23-24

The recent massive increase in capability for sequencing genomes is producing enormous advances in our understanding of biological systems. However, there is a bottleneck in genome annotation--determining the structure of all transcribed genes. Experimental data from MS studies can play a major role in confirming and correcting gene structure--proteogenomics. However, there are some technical and practical challenges to overcome, since proteogenomics requires pipelines comprising a complex set o ...[more]

PMID: 25297486

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Re-analysis of: ‘CSF proteome mapping: Human cerebrospinal fluid LC-MS/MS, Part 2’ (PXD000652) to demonstrate the ProteoAnnotator software for mapping identifications to Ensembl genomes

Project description:We have performed a re-analysis of PXD000652 - a human proteome CSF fluid sample, originally queried versus UniProt. We have used our ProteoAnnotator package to re-search the data against Ensembl Human build 76 to demonstrate the capability of the pipeline to perform genome mapping on a large scale.

2014-10-20 | PXD001390 | Pride

Genome-wide mapping of DNA re-replication in S. cerevisiae

Project description:To maintain genomic stability, re-initiation of eukaryotic DNA replication within a single cell cycle is blocked by multiple mechanisms that inactivate or remove replication proteins after G1 phase. Consistent with the prevailing notion that these mechanisms are redundant, we previously showed that simultaneous deregulation of three replication proteins, ORC, Cdc6 and Mcm2-7, was necessary to cause detectable bulk re-replication in G2/M phase in Saccharomyces cerevisiae. In this study, we used microarray comparative genomic hybridization (CGH) to provide a more comprehensive and detailed analysis of re-replication. This genome-wide analysis suggests that re-initiation in G2/M phase primarily occurs at a subset of both active and latent origins, but is independent of chromosomal determinants that specify the use and timing of these origins in S phase. We demonstrate that re-replication can be induced within S phase, but differs in amount and location fr om re-replication in G2/M phase, illustrating the dynamic nature of DNA replication controls. Finally, we show that very limited re-replication can be detected by microarray CGH when only two replication proteins are deregulated, suggesting that the mechanisms blocking re-replication are not redundant. Therefore we propose that eukaryotic re-replication at levels below current detection limits may be more prevalent and a greater source of genomic instability than previously appreciated. Keywords: comparative genomic hybridization (CGH), DNA replication, re-replication

2006-03-17 | GSE4181 | GEO

Relative Retention Time Estimation Improves N-Glycopeptide Identifications by LC-MS/MS.

Project description: Not available

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REPARATION: Ribosome Profiling Assisted (Re-) Annotation of Bacterial genomes

Project description:Prokaryotic genome annotation is highly dependent on automated methods, as manual curation cannot keep up with the exponential growth of sequenced genomes. Current automated techniques depend heavily on sequence context and often underestimate the complexity of the proteome. We developed REPARATION (RibosomeE Profiling Assisted (Re-)AnnotaTION), a de novo algorithm that takes advantage of experimental evidence from ribosome profiling (Ribo-seq) to delineate translated open reading frames (ORFs) in bacteria, independent of genome annotation. Ribo-seq next generation sequencing technique that provides a genome-wide snapshot of the position translating ribosome along an mRNA at the time of the experiment. REPARATION evaluates all possible ORFs in the genome and estimates minimum thresholds to screen for spurious ORFs based on a growth curve model. We applied REPARATION to three annotated bacterial species to obtain a more comprehensive mapping of their translation landscape in support of experimental data. In all cases, we identified hundreds of novel ORFs including variants of previously annotated and novel small ORFs (<71 codons). Our predictions were supported by matching mass spectrometry (MS) proteomics data and sequence conservation analysis. REPARATION is unique in that it makes use of experimental Ribo-seq data to perform de novo ORF delineation in bacterial genomes, and thus can identify putative coding ORFs irrespective of the sequence context of the reading frame.

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Re-sequencing and optical mapping reveals misassemblies and real inversions on Corynebacterium pseudotuberculosis genomes.

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REPARATION: Ribosome Profiling Assisted (Re-) Annotation of Bacterial genomes

Project description:The delineation of genes in bacteria has remained an important challenge because prokaryotic genomes are often tightly packed frequently resulting in overlapping genes. We hereby present a de novo approach called REPARATION (RibosomeE Profiling Assisted (Re-)AnnotaTION) to delineate translated open reading frames (ORFs) in bacteria independent of (available) genome annotation. By deep sequencing of ribosome protected mRNA fragments (RPF) to map translating ribosomes across the entire genome, REPARATION takes advantage of the recently developed ribosome profiling (Ribo-seq) technique. REPARATION starts by traversing the entire genome to generate all possible ORFs and then collects their corresponding RPF signal information. Based on a growth curve model to estimate minimum ORF read density and Ribo-seq RPF coverage, thresholds indicative of translation is estimated. Finally, our algorithm applies a random forest model to build a classifier to classify putative protein coding ORFs. We evaluated the performance of REPARATION on 3 annotated bacterial species using in-house generated Ribo-seq data and matching N-terminal and shotgun proteomics data next to publically available Ribo-seq data. In all cases, about 80% of the ORFs predicted by REPARATION were previously annotated as protein coding. While 13-20% were variants of previously annotated ORFs and about 3-4% point to novel translated ORFs within intergenic or other regions previously annotated as non-coding. Without stringent length restrictions REPARATION was able to identify several small ORFs (sORFs). Multiple supportive evidence from matching MS data and sequence conservation analysis was obtained to validate predicted ORFs.

2017-01-30 | GSE91066 | GEO

Improved application of RNAModMapper - An RNA modification mapping software tool - For analysis of liquid chromatography tandem mass spectrometry (LC-MS/MS) data.

Project description: Not available

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