2024-03-28T13:45:22Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/623892021-11-22T12:45:39Zcom_10261_86com_10261_1col_10261_339
DIGITAL.CSIC
author
Giraldo, R.
2012-12-11T11:33:34Z
2012-12-11T11:33:34Z
2007-10-30
Proceedings of the National Academy of Sciences, 104 (44) : 17388-17393 (2007)
1091-6490
http://hdl.handle.net/10261/62389
10.1073/pnas.0702006104
17959784
RepA, the replication initiator protein of Pseudomonas pPS10 plasmid, is made of two winged-helix (WH) domains. RepA dimers undergo a structural transformation upon binding to origin DNA sequences (iterons), resulting in monomerization and α-helix into β-strand conversion. This affects the N-terminal domain (WH1) and generates a metastable intermediate. Here it is shown that the interaction of short dsDNA oligonucleotides, including iteron or operator RepA targets, with the isolated WH1 domain promotes the assembly of different nanostructures. These range from irregular aggregates to amyloid spheroids and fibers. Their intrinsic order inversely correlates with the extent of the transformation induced by each DNA sequence on RepA. However, DNA is not a constituent of the assembled fibers, in agreement with the protein-only principle for amyloid structure. Thus, the RepA-WH1 domain on DNA binding mimics the behavior of the mammalian prion protein. The stretch of amino acids responsible for WH1 aggregation has been identified, leading to the design of mutants with enhanced or reduced amyloidogenicity and the synthesis of a peptide that assembles into a cross-β structure. RepA amyloid assemblies could have a role in the negative regulation of plasmid replication. This article underlines the potential of specific nucleic acid sequences in promoting protein amyloidogenesis at nearly physiological conditions
eng
closedAccess
Amyloidogenesis
Conformational changes
DNA binding
fiber assembly
RepA protein
Defined DNA sequences promote the assembly of a bacterial protein into distinct amyloid nanostructures
artículo
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URL
https://digital.csic.es/bitstream/10261/62389/1/restringido.pdf
File
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URL
https://digital.csic.es/bitstream/10261/62389/4/restringido.pdf.txt
File
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restringido.pdf.txt