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231225s2018 xx |||||o 00| ||eng c |
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|a 10.1021/acs.langmuir.8b01812
|2 doi
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|a pubmed24n0956.xml
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|a (DE-627)NLM286843072
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|a (NLM)30044093
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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Schaub, Jeffrey M
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Assessing Protein Dynamics on Low-Complexity Single-Stranded DNA Curtains
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1 |
|c 2018
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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| 500 |
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|a Date Completed 05.08.2019
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|a Date Revised 25.02.2020
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a Single-stranded DNA (ssDNA) is a critical intermediate in all DNA transactions. Because ssDNA is more flexible than double-stranded (ds) DNA, interactions with ssDNA-binding proteins (SSBs) may significantly compact or elongate the ssDNA molecule. Here, we develop and characterize low-complexity ssDNA curtains, a high-throughput single-molecule assay to simultaneously monitor protein binding and correlated ssDNA length changes on supported lipid bilayers. Low-complexity ssDNA is generated via rolling circle replication of short synthetic oligonucleotides, permitting control over the sequence composition and secondary structure-forming propensity. One end of the ssDNA is functionalized with a biotin, while the second is fluorescently labeled to track the overall DNA length. Arrays of ssDNA molecules are organized at microfabricated barriers for high-throughput single-molecule imaging. Using this assay, we demonstrate that E. coli SSB drastically and reversibly compacts ssDNA templates upon changes in NaCl concentration. We also examine the interactions between a phosphomimetic RPA and ssDNA. Our results indicate that RPA-ssDNA interactions are not significantly altered by these modifications. We anticipate that low-complexity ssDNA curtains will be broadly useful for single-molecule studies of ssDNA-binding proteins involved in DNA replication, transcription, and repair
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| 650 |
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|a Journal Article
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| 650 |
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|a Research Support, N.I.H., Extramural
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| 650 |
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4 |
|a Research Support, Non-U.S. Gov't
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| 650 |
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4 |
|a Research Support, U.S. Gov't, Non-P.H.S.
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| 650 |
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7 |
|a DNA, Single-Stranded
|2 NLM
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| 650 |
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7 |
|a DNA-Binding Proteins
|2 NLM
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| 650 |
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|a Escherichia coli Proteins
|2 NLM
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| 650 |
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|a RPA1 protein, human
|2 NLM
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| 650 |
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7 |
|a Replication Protein A
|2 NLM
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| 650 |
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|a SSB protein, E coli
|2 NLM
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| 650 |
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|a Green Fluorescent Proteins
|2 NLM
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| 650 |
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7 |
|a 147336-22-9
|2 NLM
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| 650 |
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7 |
|a Sodium Chloride
|2 NLM
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| 650 |
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7 |
|a 451W47IQ8X
|2 NLM
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| 650 |
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|a DNA-Directed DNA Polymerase
|2 NLM
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| 650 |
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7 |
|a EC 2.7.7.7
|2 NLM
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| 700 |
1 |
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|a Zhang, Hongshan
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Soniat, Michael M
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Finkelstein, Ilya J
|e verfasserin
|4 aut
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| 773 |
0 |
8 |
|i Enthalten in
|t Langmuir : the ACS journal of surfaces and colloids
|d 1992
|g 34(2018), 49 vom: 11. Dez., Seite 14882-14890
|w (DE-627)NLM098181009
|x 1520-5827
|7 nnns
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| 773 |
1 |
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|g volume:34
|g year:2018
|g number:49
|g day:11
|g month:12
|g pages:14882-14890
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| 856 |
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|u http://dx.doi.org/10.1021/acs.langmuir.8b01812
|3 Volltext
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|h 14882-14890
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