Structural basis of ubiquitin recognition by translesion synthesis DNA polymerase ℓ

Gaofeng Cui, Robert C. Benirschke, Han Fang Tuan, Nenad Juranić, Slobodan I Macura, Maria Victoria Botuyan, Georges Mer

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Cells have evolved mutagenic bypass mechanisms that prevent stalling of the replication machinery at DNA lesions. This process, translesion DNA synthesis (TLS), involves switching from high-fidelity DNA polymerases to specialized DNA polymerases that replicate through a variety of DNA lesions. In eukaryotes, polymerase switching during TLS is regulated by the DNA damage-triggered monoubiquitylation of PCNA. How the switch operates is unknown, but all TLS polymerases of the so-called Y-family possess PCNA and ubiquitin-binding domains that are important for their function. To gain insight into the structural mechanisms underlying the regulation of TLS by ubiquitylation, we have probed the interaction of ubiquitin with a conserved ubiquitin-binding motif (UBM2) of Y-family polymerase Polℓ. Using NMR spectroscopy, we have determined the structure of a complex of human Polℓ UBM2 and ubiquitin, revealing a novel ubiquitin recognition fold consisting of two α-helices separated by a central trans-proline residue conserved in all UBMs. We show that, different from the majority of ubiquitin complexes characterized to date, ubiquitin residue Ile44 only plays a modest role in the association of ubiquitin with Polℓ UBM2. Instead, binding of UBM2 is centered on the recognition of Leu8 in ubiquitin, which is essential for the interaction.

Original languageEnglish (US)
Pages (from-to)10198-10207
Number of pages10
JournalBiochemistry
Volume49
Issue number47
DOIs
StatePublished - Nov 30 2010

Fingerprint

DNA-Directed DNA Polymerase
Ubiquitin
DNA
Proliferating Cell Nuclear Antigen
Ubiquitination
Eukaryota
Proline
Nuclear magnetic resonance spectroscopy
DNA Damage
Machinery
Magnetic Resonance Spectroscopy
Switches
Association reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structural basis of ubiquitin recognition by translesion synthesis DNA polymerase ℓ. / Cui, Gaofeng; Benirschke, Robert C.; Tuan, Han Fang; Juranić, Nenad; Macura, Slobodan I; Botuyan, Maria Victoria; Mer, Georges.

In: Biochemistry, Vol. 49, No. 47, 30.11.2010, p. 10198-10207.

Research output: Contribution to journalArticle

Cui, G, Benirschke, RC, Tuan, HF, Juranić, N, Macura, SI, Botuyan, MV & Mer, G 2010, 'Structural basis of ubiquitin recognition by translesion synthesis DNA polymerase ℓ', Biochemistry, vol. 49, no. 47, pp. 10198-10207. https://doi.org/10.1021/bi101303t
Cui G, Benirschke RC, Tuan HF, Juranić N, Macura SI, Botuyan MV et al. Structural basis of ubiquitin recognition by translesion synthesis DNA polymerase ℓ. Biochemistry. 2010 Nov 30;49(47):10198-10207. https://doi.org/10.1021/bi101303t
Cui, Gaofeng ; Benirschke, Robert C. ; Tuan, Han Fang ; Juranić, Nenad ; Macura, Slobodan I ; Botuyan, Maria Victoria ; Mer, Georges. / Structural basis of ubiquitin recognition by translesion synthesis DNA polymerase ℓ. In: Biochemistry. 2010 ; Vol. 49, No. 47. pp. 10198-10207.
@article{1bafc6f1469c4dc4a549ef500f694457,
title = "Structural basis of ubiquitin recognition by translesion synthesis DNA polymerase ℓ",
abstract = "Cells have evolved mutagenic bypass mechanisms that prevent stalling of the replication machinery at DNA lesions. This process, translesion DNA synthesis (TLS), involves switching from high-fidelity DNA polymerases to specialized DNA polymerases that replicate through a variety of DNA lesions. In eukaryotes, polymerase switching during TLS is regulated by the DNA damage-triggered monoubiquitylation of PCNA. How the switch operates is unknown, but all TLS polymerases of the so-called Y-family possess PCNA and ubiquitin-binding domains that are important for their function. To gain insight into the structural mechanisms underlying the regulation of TLS by ubiquitylation, we have probed the interaction of ubiquitin with a conserved ubiquitin-binding motif (UBM2) of Y-family polymerase Polℓ. Using NMR spectroscopy, we have determined the structure of a complex of human Polℓ UBM2 and ubiquitin, revealing a novel ubiquitin recognition fold consisting of two α-helices separated by a central trans-proline residue conserved in all UBMs. We show that, different from the majority of ubiquitin complexes characterized to date, ubiquitin residue Ile44 only plays a modest role in the association of ubiquitin with Polℓ UBM2. Instead, binding of UBM2 is centered on the recognition of Leu8 in ubiquitin, which is essential for the interaction.",
author = "Gaofeng Cui and Benirschke, {Robert C.} and Tuan, {Han Fang} and Nenad Juranić and Macura, {Slobodan I} and Botuyan, {Maria Victoria} and Georges Mer",
year = "2010",
month = "11",
day = "30",
doi = "10.1021/bi101303t",
language = "English (US)",
volume = "49",
pages = "10198--10207",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "47",

}

TY - JOUR

T1 - Structural basis of ubiquitin recognition by translesion synthesis DNA polymerase ℓ

AU - Cui, Gaofeng

AU - Benirschke, Robert C.

AU - Tuan, Han Fang

AU - Juranić, Nenad

AU - Macura, Slobodan I

AU - Botuyan, Maria Victoria

AU - Mer, Georges

PY - 2010/11/30

Y1 - 2010/11/30

N2 - Cells have evolved mutagenic bypass mechanisms that prevent stalling of the replication machinery at DNA lesions. This process, translesion DNA synthesis (TLS), involves switching from high-fidelity DNA polymerases to specialized DNA polymerases that replicate through a variety of DNA lesions. In eukaryotes, polymerase switching during TLS is regulated by the DNA damage-triggered monoubiquitylation of PCNA. How the switch operates is unknown, but all TLS polymerases of the so-called Y-family possess PCNA and ubiquitin-binding domains that are important for their function. To gain insight into the structural mechanisms underlying the regulation of TLS by ubiquitylation, we have probed the interaction of ubiquitin with a conserved ubiquitin-binding motif (UBM2) of Y-family polymerase Polℓ. Using NMR spectroscopy, we have determined the structure of a complex of human Polℓ UBM2 and ubiquitin, revealing a novel ubiquitin recognition fold consisting of two α-helices separated by a central trans-proline residue conserved in all UBMs. We show that, different from the majority of ubiquitin complexes characterized to date, ubiquitin residue Ile44 only plays a modest role in the association of ubiquitin with Polℓ UBM2. Instead, binding of UBM2 is centered on the recognition of Leu8 in ubiquitin, which is essential for the interaction.

AB - Cells have evolved mutagenic bypass mechanisms that prevent stalling of the replication machinery at DNA lesions. This process, translesion DNA synthesis (TLS), involves switching from high-fidelity DNA polymerases to specialized DNA polymerases that replicate through a variety of DNA lesions. In eukaryotes, polymerase switching during TLS is regulated by the DNA damage-triggered monoubiquitylation of PCNA. How the switch operates is unknown, but all TLS polymerases of the so-called Y-family possess PCNA and ubiquitin-binding domains that are important for their function. To gain insight into the structural mechanisms underlying the regulation of TLS by ubiquitylation, we have probed the interaction of ubiquitin with a conserved ubiquitin-binding motif (UBM2) of Y-family polymerase Polℓ. Using NMR spectroscopy, we have determined the structure of a complex of human Polℓ UBM2 and ubiquitin, revealing a novel ubiquitin recognition fold consisting of two α-helices separated by a central trans-proline residue conserved in all UBMs. We show that, different from the majority of ubiquitin complexes characterized to date, ubiquitin residue Ile44 only plays a modest role in the association of ubiquitin with Polℓ UBM2. Instead, binding of UBM2 is centered on the recognition of Leu8 in ubiquitin, which is essential for the interaction.

UR - http://www.scopus.com/inward/record.url?scp=78649501997&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78649501997&partnerID=8YFLogxK

U2 - 10.1021/bi101303t

DO - 10.1021/bi101303t

M3 - Article

C2 - 21049971

AN - SCOPUS:78649501997

VL - 49

SP - 10198

EP - 10207

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 47

ER -