ZFIN ID: ZDB-PUB-140728-11
Annotation of the zebrafish genome through an integrated transcriptomic and proteomic analysis
Kelkar, D.S., Provost, E., Chaerkady, R., Muthusamy, B., Manda, S.S., Subbannayya, T., Selvan, L.D., Wang, C.H., Datta, K.K., Woo, S., Dwivedi, S.B., Renuse, S., Getnet, D., Huang, T.C., Kim, M.S., Pinto, S.M., Mitchell, C.J., Madugundu, A.K., Kumar, P., Sharma, J., Advani, J., Dey, G., Balakrishnan, L., Syed, N., Nanjappa, V., Subbannayya, Y., Goel, R., Prasad, T.S., Bafna, V., Sirdeshmukh, R., Gowda, H., Wang, C., Leach, S.D., Pandey, A.
Date: 2014
Source: Molecular & cellular proteomics : MCP   13(11): 3184-98 (Journal)
Registered Authors: Leach, Steven D., Wang, Chieh-Huei
Keywords: Gene Expression*, Omics, PCR, Proteogenomics, Transcription*
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Gene Expression Profiling
  • Genome/genetics*
  • High-Throughput Nucleotide Sequencing
  • Mass Spectrometry
  • Molecular Sequence Annotation
  • Proteome/analysis*
  • Proteome/genetics*
  • Proteomics
  • Sequence Analysis, RNA
  • Transcriptome/genetics*
  • Zebrafish/genetics*
PubMed: 25060758 Full text @ Mol. Cell. Proteomics
ABSTRACT
Accurate annotation of protein-coding genes is one of the primary tasks upon completion of whole genome sequencing of any organism. In this study, we used an integrated transcriptomic and proteomic strategy to validate and improve the existing zebrafish genome annotation. We undertook high resolution mass spectrometry-based proteomic profiling of ten adult organs, whole adult fish body and two developmental stages of zebrafish (SAT line) in addition to transcriptomic profiling of six organs. More than 7,000 proteins were identified from proteomic analyses and ~69,000 high confidence transcripts were assembled from the RNA-Seq data. Approximately 15% of the transcripts mapped to intergenic regions, the majority of which are likely long non-coding RNAs. These high quality transcriptomic and proteomic data were used to manually re-annotate the zebrafish genome. We report identification of 157 novel protein-coding genes. In addition, our data led to modification of existing gene structures which include novel exons, change in exon coordinates, change in frame of translation, translation in annotated UTRs and joining of genes. Finally, we discovered four instances of genome assembly errors that were supported by both proteomic and transcriptomic data. Our study shows how an integrative analysis of the transcriptome and the proteome can extend our understanding of even well-annotated genomes.
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