Abstract

Cold-shock domain containing protein C2 (CSD-C2; also known as PIPPin) is an RNA-binding protein conserved in the evolution that interacts with the3’-untranslated region (3’-UTR) of rat H1.0 and H3.3 histone messengers. Biolayer interferometry (BLI) is a technique that measures changes in an interferencepattern generated from visible light, reflected from an optical layer, and a biolayer which contains molecules of interest. In this study, we used the BLI methodology inorder to analyze and describe the binding properties of CSD-C2 and the mRNA encoding the rat brain histone protein H3.3. Recombinant CSD-C2 was incubatedwith in vitro transcribed, and biotinylated H3.3 RNA fragments bound to streptavidin-conjugated Octet optical biosensors. In order to define the RNA regioninvolved in binding, we used RNA probes corresponding to different portions of H3.3 RNA 3’-UTR.In this study, we showed that CSD-C2 binds to the last 199 nucleotides of the H3.3 RNA 3’-UTR, and that the apparent affinity constant of the interaction is in thenanomolar range. In addition, this study confirmed that BLI can be a very efficient and reliable method for studying RNA-protein interactions.
Lingua originaleEnglish
pagine (da-a)1-4
Numero di pagine4
RivistaBRAIN AND NERVES
Volume1
Stato di pubblicazionePublished - 2017

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Interferometry
RNA
3' Untranslated Regions
Histones
Untranslated Regions
RNA Probes
RNA-Binding Proteins
Streptavidin
Biosensing Techniques
Shock
Proteins
Nucleotides
Light
Messenger RNA
Brain

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title = "Kinetics of rat CSD-C2 binding to H3.3 RNA",
abstract = "Cold-shock domain containing protein C2 (CSD-C2; also known as PIPPin) is an RNA-binding protein conserved in the evolution that interacts with the3’-untranslated region (3’-UTR) of rat H1.0 and H3.3 histone messengers. Biolayer interferometry (BLI) is a technique that measures changes in an interferencepattern generated from visible light, reflected from an optical layer, and a biolayer which contains molecules of interest. In this study, we used the BLI methodology inorder to analyze and describe the binding properties of CSD-C2 and the mRNA encoding the rat brain histone protein H3.3. Recombinant CSD-C2 was incubatedwith in vitro transcribed, and biotinylated H3.3 RNA fragments bound to streptavidin-conjugated Octet optical biosensors. In order to define the RNA regioninvolved in binding, we used RNA probes corresponding to different portions of H3.3 RNA 3’-UTR.In this study, we showed that CSD-C2 binds to the last 199 nucleotides of the H3.3 RNA 3’-UTR, and that the apparent affinity constant of the interaction is in thenanomolar range. In addition, this study confirmed that BLI can be a very efficient and reliable method for studying RNA-protein interactions.",
author = "{Di Liegro}, Italia and Patrizia Saladino and {Di Liegro}, {Carlo Maria} and Gabriella Schiera and Daniel Gygax",
year = "2017",
language = "English",
volume = "1",
pages = "1--4",
journal = "BRAIN AND NERVES",
issn = "2515-012X",

}

TY - JOUR

T1 - Kinetics of rat CSD-C2 binding to H3.3 RNA

AU - Di Liegro, Italia

AU - Saladino, Patrizia

AU - Di Liegro, Carlo Maria

AU - Schiera, Gabriella

AU - Gygax, Daniel

PY - 2017

Y1 - 2017

N2 - Cold-shock domain containing protein C2 (CSD-C2; also known as PIPPin) is an RNA-binding protein conserved in the evolution that interacts with the3’-untranslated region (3’-UTR) of rat H1.0 and H3.3 histone messengers. Biolayer interferometry (BLI) is a technique that measures changes in an interferencepattern generated from visible light, reflected from an optical layer, and a biolayer which contains molecules of interest. In this study, we used the BLI methodology inorder to analyze and describe the binding properties of CSD-C2 and the mRNA encoding the rat brain histone protein H3.3. Recombinant CSD-C2 was incubatedwith in vitro transcribed, and biotinylated H3.3 RNA fragments bound to streptavidin-conjugated Octet optical biosensors. In order to define the RNA regioninvolved in binding, we used RNA probes corresponding to different portions of H3.3 RNA 3’-UTR.In this study, we showed that CSD-C2 binds to the last 199 nucleotides of the H3.3 RNA 3’-UTR, and that the apparent affinity constant of the interaction is in thenanomolar range. In addition, this study confirmed that BLI can be a very efficient and reliable method for studying RNA-protein interactions.

AB - Cold-shock domain containing protein C2 (CSD-C2; also known as PIPPin) is an RNA-binding protein conserved in the evolution that interacts with the3’-untranslated region (3’-UTR) of rat H1.0 and H3.3 histone messengers. Biolayer interferometry (BLI) is a technique that measures changes in an interferencepattern generated from visible light, reflected from an optical layer, and a biolayer which contains molecules of interest. In this study, we used the BLI methodology inorder to analyze and describe the binding properties of CSD-C2 and the mRNA encoding the rat brain histone protein H3.3. Recombinant CSD-C2 was incubatedwith in vitro transcribed, and biotinylated H3.3 RNA fragments bound to streptavidin-conjugated Octet optical biosensors. In order to define the RNA regioninvolved in binding, we used RNA probes corresponding to different portions of H3.3 RNA 3’-UTR.In this study, we showed that CSD-C2 binds to the last 199 nucleotides of the H3.3 RNA 3’-UTR, and that the apparent affinity constant of the interaction is in thenanomolar range. In addition, this study confirmed that BLI can be a very efficient and reliable method for studying RNA-protein interactions.

UR - http://hdl.handle.net/10447/249034

UR - http://www.oatext.com/pdf/JBN-1-104.pdf

M3 - Article

VL - 1

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EP - 4

JO - BRAIN AND NERVES

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SN - 2515-012X

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