Secretion trap tagging of secreted and membrane-spanning proteins using Arabidopsis gene traps

Groover, A. T., Fontana, J. R., Arroyo, J. M., Yordan, C., McCombie, W. R., Martienssen, R. A. (2003) Secretion trap tagging of secreted and membrane-spanning proteins using Arabidopsis gene traps. Plant Physiology, 132 (2). pp. 698-708. ISSN 00320889 (ISSN)

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URL: http://www.ncbi.nlm.nih.gov/pubmed/12805598
DOI: 10.1104/pp.103.020099

Abstract

Secreted and membrane-spanning proteins play fundamental roles in plant development but pose challenges for genetic identification and characterization. We describe a "secretion trap" screen for gene trap insertions in genes encoding proteins routed through the secretory pathway. The gene trap transposon encodes a β-glucuronidase reporter enzyme that is inhibited by N-linked glycosylation specific to the secretory pathway. Treatment of seedlings with tunicamycin inhibits glycosylation, resulting in increased activity of secreted β-glucuronidase fusions that result from gene trap integration downstream of exons encoding signal peptides. In the 2,059 gene trap lines that we screened, 32 secretion trap expression patterns were identified in a wide variety of tissues including embryos, meristems, and the developing vasculature. Genes disrupted by the secretion traps encode putative extracellular signaling proteins, membrane transport proteins, and novel secreted proteins of unknown function missed by conventional mutagenesis and gene prediction. Secretion traps provide a unique reagent for gene expression studies and can guide the genetic combination of loss of function alleles in related genes.

Item Type: Paper
Uncontrolled Keywords: Genes Membranes Mutagenesis Proteins Tissue Membrane-spanning proteins Plants (botany) Arabidopsis Arabidopsis Proteins Cell Membrane Genes, Plant Mutagenesis, Insertional Plant Roots Reverse Transcriptase Polymerase Chain Reaction Signal Transduction Embryophyta insertion sequences transposons
Subjects: organism description > plant > Arabidopsis
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics
organism description > plant
CSHL Authors:
Communities: CSHL labs > Martienssen lab
CSHL labs > McCombie lab
Depositing User: Matt Covey
Date Deposited: 23 Apr 2013 16:58
Last Modified: 23 Apr 2013 16:58
PMCID: PMC167008
Related URLs:
URI: http://repository.cshl.edu/id/eprint/28240

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