Abstract
The endoplasmic reticulum (ER) is a dynamic organelle that is amenable to major restructuring. Introduction of recombinant ER-membrane-resident proteins that form homo oligomers is a known method of inducing ER proliferation: interaction of the proteins with each other alters the local structure of the ER network, leading to the formation large aggregations of expanded ER, sometimes leading to the formation of organized smooth endoplasmic reticulum (OSER). However, these membrane structures formed by ER proliferation are poorly characterized and this hampers their potential development for plant synthetic biology. Here, we characterize a range of ER-derived membranous compartments in tobacco and show how the nature of the polyproteins introduced into the ER membrane affect the morphology of the final compartment. We show that a cytosol-facing oligomerization domain is an essential component for compartment formation. Using fluorescence recovery after photobleaching, we demonstrate that although the compartment retains a connection to the ER, a diffusional barrier exists to both the ER and the cytosol associated with the compartment. Using quantitative image analysis, we also show that the presence of the compartment does not disrupt the rest of the ER network. Moreover, we demonstrate that it is possible to recruit a heterologous, bacterial enzyme to the compartment, and for the enzyme to accumulate to high levels. Finally, transgenic Arabidopsis constitutively expressing the compartment-forming polyproteins grew and developed normally under standard conditions.
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Claims 8
Claim · model-written summary
Polyprotein nature affected morphology of ER-derived compartments in tobacco.
- Taxon
- tobacco
Nicotiana tabacume0 - Treatment
- polyproteins introduced into the ER membranee0
- Outcome
- morphology of the final compartmente0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we characterize a range of ER-derived membranous compartments in tobacco and show how the nature of the polyproteins introduced into the ER membrane affect the morphology of the final compartment.”
Claim · model-written summary
A cytosol-facing oligomerization domain was essential for compartment formation in tobacco.
- Taxon
- tobaccoNicotiana tabacume1
- Gene
- cytosol-facing oligomerization domaine0
- Outcome
- compartment formatione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a cytosol-facing oligomerization domain is an essential component for compartment formation.”
e1“in tobacco”
Claim · model-written summary
The compartment remained connected to the ER in tobacco.
- Taxon
- tobaccoNicotiana tabacume1
- Outcome
- connection to the ERe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the compartment retains a connection to the ER”
e1“in tobacco”
Claim · model-written summary
A diffusional barrier existed between the compartment and both ER and cytosol in tobacco.
- Taxon
- tobaccoNicotiana tabacume1
- Outcome
- diffusional barriere0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a diffusional barrier exists to both the ER and the cytosol associated with the compartment.”
e1“in tobacco”
Claim · model-written summary
The compartment did not disrupt the rest of the ER network in tobacco.
- Taxon
- tobaccoNicotiana tabacume1
- Outcome
- rest of the ER networke0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the presence of the compartment does not disrupt the rest of the ER network.”
e1“in tobacco”
Claim · model-written summary
A heterologous bacterial enzyme could be recruited to the compartment in tobacco.
- Taxon
- tobaccoNicotiana tabacume1
- Gene
- bacterial enzymee0
- Outcome
- recruit a heterologous, bacterial enzyme to the compartmente0
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“it is possible to recruit a heterologous, bacterial enzyme to the compartment”
e1“in tobacco”
Claim · model-written summary
The recruited bacterial enzyme accumulated to high levels in the compartment.
- Taxon
- tobaccoNicotiana tabacume2
- Gene
- bacterial enzymee1
- Outcome
- enzyme to accumulate to high levelse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“for the enzyme to accumulate to high levels.”
e1“a heterologous, bacterial enzyme to the compartment”
e2“in tobacco”
Claim · model-written summary
Transgenic Arabidopsis expressing the compartment-forming polyproteins grew and developed normally.
- Taxon
- Arabidopsis
Arabidopsise0 - Material
- transgenic Arabidopsise0
- Gene
- compartment-forming polyproteinse0
- Outcome
- grew and developed normallye0
- Qualifier
- standard conditionse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“transgenic Arabidopsis constitutively expressing the compartment-forming polyproteins grew and developed normally under standard conditions.”