Identification of new mutations at the PCNA subunit interface that block translesion synthesis

Christine M. Kondratick, Elizabeth M. Boehm, Lynne Dieckman, Kyle T. Powers, Julio C. Sanchez, Samuel R. Mueting, M. Todd Washington

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Proliferating cell nuclear antigen (PCNA) plays an essential role in DNA replication and repair by interacting with a large number of proteins involved in these processes. Two amino acid substitutions in PCNA, both located at the subunit interface, have previously been shown to block translesion synthesis (TLS), a pathway for bypassing DNA damage during replication. To better understand the role of the subunit interface in TLS, we used random mutagenesis to generate a set of 33 PCNA mutants with substitutions at the subunit interface. We assayed the full set of mutants for viability and sensitivity to ultraviolet (UV) radiation. We then selected a subset of 17 mutants and measured their rates of cell growth, spontaneous mutagenesis, and UV-induced mutagenesis. All except three of these 17 mutants were partially or completely defective in induced mutagenesis, which indicates a partial or complete loss of TLS. These results demonstrate that the integrity of the subunit interface of PCNA is essential for efficient TLS and that even conservative substitutions have the potential to disrupt this process.

Original languageEnglish
Article numbere0157023
JournalPLoS One
Volume11
Issue number6
DOIs
StatePublished - Jun 1 2016

Fingerprint

proliferating cell nuclear antigen
Mutagenesis
Proliferating Cell Nuclear Antigen
mutagenesis
mutation
mutants
Mutation
Substitution reactions
synthesis
DNA
Cell growth
amino acid substitution
DNA replication
Amino Acid Substitution
DNA repair
DNA Replication
DNA Repair
Ultraviolet radiation
DNA damage
DNA Damage

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Kondratick, C. M., Boehm, E. M., Dieckman, L., Powers, K. T., Sanchez, J. C., Mueting, S. R., & Todd Washington, M. (2016). Identification of new mutations at the PCNA subunit interface that block translesion synthesis. PLoS One, 11(6), [e0157023]. https://doi.org/10.1371/journal.pone.0157023

Identification of new mutations at the PCNA subunit interface that block translesion synthesis. / Kondratick, Christine M.; Boehm, Elizabeth M.; Dieckman, Lynne; Powers, Kyle T.; Sanchez, Julio C.; Mueting, Samuel R.; Todd Washington, M.

In: PLoS One, Vol. 11, No. 6, e0157023, 01.06.2016.

Research output: Contribution to journalArticle

Kondratick, CM, Boehm, EM, Dieckman, L, Powers, KT, Sanchez, JC, Mueting, SR & Todd Washington, M 2016, 'Identification of new mutations at the PCNA subunit interface that block translesion synthesis', PLoS One, vol. 11, no. 6, e0157023. https://doi.org/10.1371/journal.pone.0157023
Kondratick, Christine M. ; Boehm, Elizabeth M. ; Dieckman, Lynne ; Powers, Kyle T. ; Sanchez, Julio C. ; Mueting, Samuel R. ; Todd Washington, M. / Identification of new mutations at the PCNA subunit interface that block translesion synthesis. In: PLoS One. 2016 ; Vol. 11, No. 6.
@article{cda31db4c0a24ca88937e4673ed1eb69,
title = "Identification of new mutations at the PCNA subunit interface that block translesion synthesis",
abstract = "Proliferating cell nuclear antigen (PCNA) plays an essential role in DNA replication and repair by interacting with a large number of proteins involved in these processes. Two amino acid substitutions in PCNA, both located at the subunit interface, have previously been shown to block translesion synthesis (TLS), a pathway for bypassing DNA damage during replication. To better understand the role of the subunit interface in TLS, we used random mutagenesis to generate a set of 33 PCNA mutants with substitutions at the subunit interface. We assayed the full set of mutants for viability and sensitivity to ultraviolet (UV) radiation. We then selected a subset of 17 mutants and measured their rates of cell growth, spontaneous mutagenesis, and UV-induced mutagenesis. All except three of these 17 mutants were partially or completely defective in induced mutagenesis, which indicates a partial or complete loss of TLS. These results demonstrate that the integrity of the subunit interface of PCNA is essential for efficient TLS and that even conservative substitutions have the potential to disrupt this process.",
author = "Kondratick, {Christine M.} and Boehm, {Elizabeth M.} and Lynne Dieckman and Powers, {Kyle T.} and Sanchez, {Julio C.} and Mueting, {Samuel R.} and {Todd Washington}, M.",
year = "2016",
month = "6",
day = "1",
doi = "10.1371/journal.pone.0157023",
language = "English",
volume = "11",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "6",

}

TY - JOUR

T1 - Identification of new mutations at the PCNA subunit interface that block translesion synthesis

AU - Kondratick, Christine M.

AU - Boehm, Elizabeth M.

AU - Dieckman, Lynne

AU - Powers, Kyle T.

AU - Sanchez, Julio C.

AU - Mueting, Samuel R.

AU - Todd Washington, M.

PY - 2016/6/1

Y1 - 2016/6/1

N2 - Proliferating cell nuclear antigen (PCNA) plays an essential role in DNA replication and repair by interacting with a large number of proteins involved in these processes. Two amino acid substitutions in PCNA, both located at the subunit interface, have previously been shown to block translesion synthesis (TLS), a pathway for bypassing DNA damage during replication. To better understand the role of the subunit interface in TLS, we used random mutagenesis to generate a set of 33 PCNA mutants with substitutions at the subunit interface. We assayed the full set of mutants for viability and sensitivity to ultraviolet (UV) radiation. We then selected a subset of 17 mutants and measured their rates of cell growth, spontaneous mutagenesis, and UV-induced mutagenesis. All except three of these 17 mutants were partially or completely defective in induced mutagenesis, which indicates a partial or complete loss of TLS. These results demonstrate that the integrity of the subunit interface of PCNA is essential for efficient TLS and that even conservative substitutions have the potential to disrupt this process.

AB - Proliferating cell nuclear antigen (PCNA) plays an essential role in DNA replication and repair by interacting with a large number of proteins involved in these processes. Two amino acid substitutions in PCNA, both located at the subunit interface, have previously been shown to block translesion synthesis (TLS), a pathway for bypassing DNA damage during replication. To better understand the role of the subunit interface in TLS, we used random mutagenesis to generate a set of 33 PCNA mutants with substitutions at the subunit interface. We assayed the full set of mutants for viability and sensitivity to ultraviolet (UV) radiation. We then selected a subset of 17 mutants and measured their rates of cell growth, spontaneous mutagenesis, and UV-induced mutagenesis. All except three of these 17 mutants were partially or completely defective in induced mutagenesis, which indicates a partial or complete loss of TLS. These results demonstrate that the integrity of the subunit interface of PCNA is essential for efficient TLS and that even conservative substitutions have the potential to disrupt this process.

UR - http://www.scopus.com/inward/record.url?scp=84973303768&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84973303768&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0157023

DO - 10.1371/journal.pone.0157023

M3 - Article

VL - 11

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 6

M1 - e0157023

ER -