Data_Sheet_1_Agrobacterium VirE3 Uses Its Two Tandem Domains at the C-Terminus to Retain Its Companion VirE2 on the Cytoplasmic Side of the Host Plasma Membrane.docx
Agrobacterium tumefaciens is the causal agent of crown gall disease in nature; in the laboratory the bacterium is widely used for plant genetic modification. The bacterium delivers a single-stranded transferred DNA (T-DNA) and a group of crucial virulence proteins into host cells. A putative T-complex is formed inside host cells that is composed of T-DNA and virulence proteins VirD2 and VirE2, which protect the foreign DNA from degradation and guide its way into the host nucleus. However, little is known about how the T-complex is assembled inside host cells. We combined the split-GFP and split-sfCherry labeling systems to study the interaction of Agrobacterium-delivered VirE2 and VirE3 in host cells. Our results indicated that VirE2 co-localized with VirE3 on the cytoplasmic side of the host cellular membrane upon the delivery. We identified and characterized two tandem domains at the VirE3 C-terminus that interacted with VirE2 in vitro. Deletion of these two domains abolished the VirE2 accumulation on the host plasma membrane and affected the transformation. Furthermore, the two VirE2-interacting domains of VirE3 exhibited different affinities with VirE2. Collectively, this study demonstrates that the anchorage protein VirE3 uses the two tandem VirE2-interacting domains to facilitate VirE2 protection for T-DNA at the cytoplasmic side of the host cell entrance.
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References
- https://doi.org//10.1128/JB.169.3.1046-1055.1987
- https://doi.org//10.1105/tpc.108.060467
- https://doi.org//10.1002/j.1460-2075.1995.tb07323.x
- https://doi.org//10.1038/nbt1044
- https://doi.org//10.1007/s004380050432
- https://doi.org//10.1038/nrmicro753
- https://doi.org//10.1126/science.1095211
- https://doi.org//10.1146/annurev-biochem-062911-102821
- https://doi.org//10.1016/0092-8674(77)90043-5
- https://doi.org//10.1146/annurev.micro.58.030603.123630
- https://doi.org//10.1128/JB.170.6.2659-2667.1988
- https://doi.org//10.1111/j.1462-5822.2006.00830.x
- https://doi.org//10.1073/pnas.86.4.1193
- https://doi.org//10.1038/nbt0998-839
- https://doi.org//10.1073/pnas.0801525105
- https://doi.org//10.1038/s41467-017-00494-8
- https://doi.org//10.1038/s42003-019-0589-x
- https://doi.org//10.1074/jbc.M605270200
- https://doi.org//10.1093/nar/gkl877
- https://doi.org//10.1146/annurev-phyto-080508-081852
- https://doi.org//10.1105/tpc.17.00027
- https://doi.org//10.1016/S0378-1119(98)00130-9
- https://doi.org//10.1128/JB.186.10.3065-3077.2004
- https://doi.org//10.1128/JB.01018-08
- https://doi.org//10.1128/JB.00747-06
- https://doi.org//10.1199/tab.0186
- https://doi.org//10.1186/s40694-017-0035-0
- https://doi.org//10.1128/jb.182.6.1774-1778.2000
- https://doi.org//10.1038/ncomms11046
- https://doi.org//10.1111/j.1529-8817.2009.00688.x
- https://doi.org//10.1371/journal.ppat.1005502
- https://doi.org//10.1038/sj.emboj.7600524
- https://doi.org//10.1073/pnas.0404118102
- https://doi.org//10.1126/sciadv.1601528
- https://doi.org//10.1016/j.celrep.2018.09.023
- https://doi.org//10.1111/tpj.12397
- https://doi.org//10.1146/annurev.cellbio.22.011105.102022
- https://doi.org//10.1007/s00294-005-0578-0
- https://doi.org//10.1111/j.1365-313X.2007.03212.x
- https://doi.org//10.3389/fpls.2014.00730
- https://doi.org//10.1111/tpj.13048
- https://doi.org//10.1105/tpc.17.00047
- https://doi.org//10.1073/pnas.93.4.1613
- https://doi.org//10.1038/emboj.2010.8
- https://doi.org//10.1073/pnas.93.1.126
- https://doi.org//10.1002/mbo3.152
- https://doi.org//10.1074/jbc.270.3.1269
- https://doi.org//10.1093/nar/gkg179
- https://doi.org//10.1128/JB.171.5.2573-2580.1989
- https://doi.org//10.1073/pnas.87.17.6684
- https://doi.org//10.1038/318624a0
- https://doi.org//10.1002/j.1460-2075.1986.tb04381.x
- https://doi.org//10.1016/0092-8674(86)90653-7
- https://doi.org//10.1007/82_2018_131
- https://doi.org//10.1016/j.copbio.2006.01.009
- https://doi.org//10.1093/emboj/20.13.3596
- https://doi.org//10.1038/nmeth.1437
- https://doi.org//10.1126/science.290.5493.979
- https://doi.org//10.1073/pnas.0406241102
- https://doi.org//10.1016/0092-8674(84)90500-2
- https://doi.org//10.1073/pnas.1612098114
- https://doi.org//10.1016/0092-8674(86)90604-5
- https://doi.org//10.1073/pnas.91.8.2994
- https://doi.org//10.1126/science.6251546
- https://doi.org//10.1128/JB.181.14.4342-4352.1999