Refined requirements for protein regions important for activity of the TALE AvrBs3

PloS One
Tom SchreiberUlla Bonas

Abstract

AvrBs3, the archetype of the family of transcription activator-like (TAL) effectors from phytopathogenic Xanthomonas bacteria, is translocated by the type III secretion system into the plant cell. AvrBs3 localizes to the plant cell nucleus and activates the transcription of target genes. Crucial for this is the central AvrBs3 region of 17.5 34-amino acid repeats that functions as a DNA-binding domain mediating recognition in a "one-repeat-to-one base pair" manner. Although AvrBs3 forms homodimers in the plant cell cytosol prior to nuclear import, it binds DNA as a monomer. Here, we show that complex formation of AvrBs3 proteins negatively affects their DNA-binding affinity in vitro. The conserved cysteine residues at position 30 of each repeat facilitate AvrBs3 complexes via disulfide bonds in vitro but are also required for the gene-inducing activity of the AvrBs3 monomer, i.e., activation of plant gene promoters. Our data suggest that the latter is due to a contribution to protein plasticity and that cysteine substitutions to alanine or serine result in a different DNA-binding mode. In addition, our studies revealed that extended parts of both the N-terminal and C-terminal regions of AvrBs3 contribute to DNA binding and, henc...Continue Reading

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Citations

Aug 16, 2016·Nature Chemical Biology·Luke CuculisCharles M Schroeder
Sep 21, 2016·Nature Chemical Biology·Sebastian Becker, Jens Boch
Dec 15, 2016·Molecular Plant·Li WangAdam J Bogdanove
May 2, 2017·Nucleic Acids Research·Fabio C RinaldiAdam J Bogdanove
Dec 30, 2017·The Plant Journal : for Cell and Molecular Biology·Jessica L EricksonMartin H Schattat
Dec 22, 2019·Nucleic Acids Research·Luke CuculisCharles M Schroeder
May 12, 2016·Seminars in Cell & Developmental Biology·Karl J SchreiberJennifer D Lewis

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Methods Mentioned

BETA
electrophoretic mobility shift
circular dichroism
transgenic
X-ray
Electrophoresis

Software Mentioned

ClustalW
Boxshade
Kaleidagraph
Synergy
ZEN

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