Production, purification of the recombinant analog of y-box-binding protein 1 and its interaction with poly(ADP-ribose), RNA, single- and double-stranded DNAs

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Production, purification of the recombinant analog of y-box-binding protein 1 and its interaction with poly(ADP-ribose), RNA, single- and double-stranded DNAs. / Alemasova, E. E.; Naumenko, K. N.; Pestryakov, P. E. et al.

In: Biopolymers and Cell, Vol. 33, No. 3, 2017, p. 214-220.

Research output: Contribution to journal › Article › peer-review

BibTeX

@article{3e12c1683680410fb03c638dc03bc6d1,

title = "Production, purification of the recombinant analog of y-box-binding protein 1 and its interaction with poly(ADP-ribose), RNA, single- and double-stranded DNAs",

abstract = "Aim. Production and purification of the recombinant histidine-tagged Y-box- binding protein 1 and study of its interaction with DNA, RNA and poly(ADP-ribose). Methods. Ligationindependent cloning, PCR, Sanger sequencing, protein chromatography, polyacrylamide gel electrophoresis, and electrophoresis mobility shift assay. Results. cDNA coding for the YB-1 protein has a previously undocumented two single nucleotide polymorphisms. The expression construct for production of the his-tagged YB-1 protein was designed to simplify the purification procedure and an appropriate protocol for protein purification was developed. Using electrophoresis mobility shift assay, we have shown that poly(ADP-ribose) competes with a double- and single-stranded DNA and RNA for binding to purified recombinant his-tagged YB-1. Conclusions. In the present work we developed and optimized the procedure of the recombinant YB-1 protein production and purification from bacterial cells. We found that poly(ADP-ribose) at high concentration is able to recruit YB-1 protein from the YB-1-DNA and YB-1-RNA complexes, suggesting a possible YB-1 involvement in DNA repair.",

keywords = "DNA repair, Poly(ADP-ribose) (PAR), Protein purification, YB-1",

author = "Alemasova, {E. E.} and Naumenko, {K. N.} and Pestryakov, {P. E.} and Lavrik, {O. I.}",

note = "Publisher Copyright: {\textcopyright} 2017 E. E. Alemasova et al.",

year = "2017",

doi = "10.7124/bc.000954",

language = "English",

volume = "33",

pages = "214--220",

journal = "Biopolymers and Cell",

issn = "0233-7657",

publisher = "National Academy of Sciences of Ukraine",

number = "3",

}

RIS

TY - JOUR

T1 - Production, purification of the recombinant analog of y-box-binding protein 1 and its interaction with poly(ADP-ribose), RNA, single- and double-stranded DNAs

AU - Alemasova, E. E.

AU - Naumenko, K. N.

AU - Pestryakov, P. E.

AU - Lavrik, O. I.

PY - 2017

Y1 - 2017

N2 - Aim. Production and purification of the recombinant histidine-tagged Y-box- binding protein 1 and study of its interaction with DNA, RNA and poly(ADP-ribose). Methods. Ligationindependent cloning, PCR, Sanger sequencing, protein chromatography, polyacrylamide gel electrophoresis, and electrophoresis mobility shift assay. Results. cDNA coding for the YB-1 protein has a previously undocumented two single nucleotide polymorphisms. The expression construct for production of the his-tagged YB-1 protein was designed to simplify the purification procedure and an appropriate protocol for protein purification was developed. Using electrophoresis mobility shift assay, we have shown that poly(ADP-ribose) competes with a double- and single-stranded DNA and RNA for binding to purified recombinant his-tagged YB-1. Conclusions. In the present work we developed and optimized the procedure of the recombinant YB-1 protein production and purification from bacterial cells. We found that poly(ADP-ribose) at high concentration is able to recruit YB-1 protein from the YB-1-DNA and YB-1-RNA complexes, suggesting a possible YB-1 involvement in DNA repair.

AB - Aim. Production and purification of the recombinant histidine-tagged Y-box- binding protein 1 and study of its interaction with DNA, RNA and poly(ADP-ribose). Methods. Ligationindependent cloning, PCR, Sanger sequencing, protein chromatography, polyacrylamide gel electrophoresis, and electrophoresis mobility shift assay. Results. cDNA coding for the YB-1 protein has a previously undocumented two single nucleotide polymorphisms. The expression construct for production of the his-tagged YB-1 protein was designed to simplify the purification procedure and an appropriate protocol for protein purification was developed. Using electrophoresis mobility shift assay, we have shown that poly(ADP-ribose) competes with a double- and single-stranded DNA and RNA for binding to purified recombinant his-tagged YB-1. Conclusions. In the present work we developed and optimized the procedure of the recombinant YB-1 protein production and purification from bacterial cells. We found that poly(ADP-ribose) at high concentration is able to recruit YB-1 protein from the YB-1-DNA and YB-1-RNA complexes, suggesting a possible YB-1 involvement in DNA repair.

KW - DNA repair

KW - Poly(ADP-ribose) (PAR)

KW - Protein purification

KW - YB-1

UR - http://www.scopus.com/inward/record.url?scp=85028918029&partnerID=8YFLogxK

U2 - 10.7124/bc.000954

DO - 10.7124/bc.000954

M3 - Article

AN - SCOPUS:85028918029

VL - 33

SP - 214

EP - 220

JO - Biopolymers and Cell

JF - Biopolymers and Cell

SN - 0233-7657

IS - 3

ER -

ID: 8681044