Research output: Contribution to journal › Article › peer-review
Dynamic light scattering study of base excision DNA repair proteins and their complexes. / Vasil'eva, Inna A.; Anarbaev, Rashid O.; Moor, Nina A. et al.
In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1867, No. 3, 01.03.2019, p. 297-305.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Dynamic light scattering study of base excision DNA repair proteins and their complexes
AU - Vasil'eva, Inna A.
AU - Anarbaev, Rashid O.
AU - Moor, Nina A.
AU - Lavrik, Olga I.
N1 - Publisher Copyright: © 2018 Elsevier B.V.
PY - 2019/3/1
Y1 - 2019/3/1
N2 - Base excision repair (BER) involves many enzymes acting in a coordinated fashion at the most common types of DNA damage. The coordination is facilitated by interactions between the enzymes and accessory proteins, X-ray repair cross-complementing protein 1 (XRCC1) and poly(ADP-ribose) polymerase 1 (PARP1). Here we use dynamic light scattering (DLS) technique to determine the hydrodynamic sizes of several BER enzymes and proteins, DNA polymerase β (Polβ), apurinic/apyrimidinic endonuclease 1 (APE1), tyrosyl-DNA phosphodiesterase 1 (TDP1), XRCC1 and PARP1, present alone or in the equimolar mixtures with each other. From the DLS data combined with glutaraldehyde cross-linking experiments and previous quantitative binding data the oligomeric states of BER proteins and their complexes are estimated. All the proteins have been proposed to form homodimers upon their self-association. The most probable oligomerization state of the binary complexes formed by PARP1 with various proteins is a heterotetramer. The oligomerization state of the binary complexes formed by XRCC1 varies from heterodimer to heterotetramer, depending on the partner. The DLS technique is applied for the first time to measure the hydrodynamic sizes of PARP1 molecules covalently bound with poly(ADP-ribose) (PAR) synthesized upon the automodification reaction. PARP1 has been detected to form huge conglomerates stabilized by Mg2+ coordinated bonds with PAR polymers.
AB - Base excision repair (BER) involves many enzymes acting in a coordinated fashion at the most common types of DNA damage. The coordination is facilitated by interactions between the enzymes and accessory proteins, X-ray repair cross-complementing protein 1 (XRCC1) and poly(ADP-ribose) polymerase 1 (PARP1). Here we use dynamic light scattering (DLS) technique to determine the hydrodynamic sizes of several BER enzymes and proteins, DNA polymerase β (Polβ), apurinic/apyrimidinic endonuclease 1 (APE1), tyrosyl-DNA phosphodiesterase 1 (TDP1), XRCC1 and PARP1, present alone or in the equimolar mixtures with each other. From the DLS data combined with glutaraldehyde cross-linking experiments and previous quantitative binding data the oligomeric states of BER proteins and their complexes are estimated. All the proteins have been proposed to form homodimers upon their self-association. The most probable oligomerization state of the binary complexes formed by PARP1 with various proteins is a heterotetramer. The oligomerization state of the binary complexes formed by XRCC1 varies from heterodimer to heterotetramer, depending on the partner. The DLS technique is applied for the first time to measure the hydrodynamic sizes of PARP1 molecules covalently bound with poly(ADP-ribose) (PAR) synthesized upon the automodification reaction. PARP1 has been detected to form huge conglomerates stabilized by Mg2+ coordinated bonds with PAR polymers.
KW - DNA base excision repair complexes
KW - Dynamic light scattering
KW - Oligomeric state
KW - Poly(ADP-ribosyl)ation
KW - Protein-protein interactions
KW - SPECIFICITY
KW - XRCC1
KW - MECHANISMS
KW - IDENTIFICATION
KW - DAMAGE
KW - IN-VITRO
KW - POLY(ADP-RIBOSE) POLYMERASE 1
KW - BINDING
KW - ASSOCIATION
KW - DOMAINS
KW - DNA Polymerase beta/chemistry
KW - X-ray Repair Cross Complementing Protein 1/chemistry
KW - Poly (ADP-Ribose) Polymerase-1/chemistry
KW - DNA Repair
KW - Phosphoric Diester Hydrolases/chemistry
KW - Dynamic Light Scattering
KW - DNA-(Apurinic or Apyrimidinic Site) Lyase/chemistry
UR - http://www.scopus.com/inward/record.url?scp=85055124596&partnerID=8YFLogxK
U2 - 10.1016/j.bbapap.2018.10.009
DO - 10.1016/j.bbapap.2018.10.009
M3 - Article
C2 - 30321662
AN - SCOPUS:85055124596
VL - 1867
SP - 297
EP - 305
JO - Biochimica et Biophysica Acta - Proteins and Proteomics
JF - Biochimica et Biophysica Acta - Proteins and Proteomics
SN - 1570-9639
IS - 3
ER -
ID: 17171645