Integrating mRNA and Protein Sequencing Enables the Detection and Quantitative Profiling of Natural Protein Sequence Variants of Populus trichocarpa

Paul E. Abraham, Xiaojing Wang, Priya Ranjan, Intawat Nookaew, Bing Zhang, Gerald A. Tuskan, Robert L. Hettich

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Next-generation sequencing has transformed the ability to link genotypes to phenotypes and facilitates the dissection of genetic contribution to complex traits. However, it is challenging to link genetic variants with the perturbed functional effects on proteins encoded by such genes. Here we show how RNA sequencing can be exploited to construct genotype-specific protein sequence databases to assess natural variation in proteins, providing information about the molecular toolbox driving cellular processes. For this study, we used two natural genotypes selected from a recent genome-wide association study of Populus trichocarpa, an obligate outcrosser with tremendous phenotypic variation across the natural population. This strategy allowed us to comprehensively catalogue proteins containing single amino acid polymorphisms (SAAPs), as well as insertions and deletions. We profiled the frequency of 128 types of naturally occurring amino acid substitutions, including both expected (neutral) and unexpected (non-neutral) SAAPs, with a subset occurring in regions of the genome having strong polymorphism patterns consistent with recent positive and/or divergent selection. By zeroing in on the molecular signatures of these important regions that might have previously been uncharacterized, we now provide a high-resolution molecular inventory that should improve accessibility and subsequent identification of natural protein variants in future genotype-to-phenotype studies.

Original languageEnglish (US)
Pages (from-to)5318-5326
Number of pages9
JournalJournal of Proteome Research
Volume14
Issue number12
DOIs
StatePublished - Dec 4 2015
Externally publishedYes

Keywords

  • RNA sequencing
  • genotype-specific protein database
  • mass spectrometry
  • natural variants
  • shotgun proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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