Secretion Trap Tagging of Secreted and Membrane-Spanning Proteins Using Arabidopsis Gene Traps1
AUTOR(ES)
Groover, Andrew T.
FONTE
The American Society for Plant Biologists
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=167008Documentos Relacionados
- Predicting the orientation of eukaryotic membrane-spanning proteins.
- Interaction and conformational dynamics of membrane-spanning protein helices
- Disruption of an Internal Membrane-Spanning Region in Shiga Toxin 1 Reduces Cytotoxicity
- The Role of the Membrane-spanning Domain Sequence in Glycoprotein-mediated Membrane Fusion
- Efficient gene tagging in Arabidopsis thaliana using a gene trap approach