Regulation of translesion synthesis DNA polymerase eta by monoubiquitination.
DNA polymerase eta is a Y family polymerase involved in translesion synthesis (TLS). Its action is initiated by simultaneous interaction between the PIP box in pol eta and PCNA and between the UBZ in pol eta and monoubiquitin attached to PCNA. Whereas monoubiquitination of PCNA is required for its interaction with pol eta during TLS, we now show that monoubiquitination of pol eta inhibits this interaction, preventing its functions in undamaged cells. Identification of monoubiquitination sites within pol eta nuclear localization signal (NLS) led to the discovery that pol eta NLS directly contacts PCNA, forming an extended pol eta-PCNA interaction surface. We name this the PCNA-interacting region (PIR) and show that its monoubiquitination is downregulated by various DNA-damaging agents. We propose that this mechanism ensures optimal availability of nonubiquitinated, TLS-competent pol eta after DNA damage. Our work shows how monoubiquitination can either positively or negatively regulate the assembly of a protein complex, depending on which substrates are targeted by ubiquitin.
Bienko
Marzena
M
Institute of Biochemistry II, Goethe University Medical School, Theodor-Stern-Kai 7, D-60590 Frankfurt am Main, Germany.
Green
Catherine M
CM
Sabbioneda
Simone
S
Crosetto
Nicola
N
Matic
Ivan
I
Hibbert
Richard G
RG
Begovic
Tihana
T
Niimi
Atsuko
A
Mann
Matthias
M
Lehmann
Alan R
AR
Dikic
Ivan
I
D000595:Amino Acid Sequence; D002460:Cell Line; D004249:DNA Damage; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D009153:Mutagens / Q000494:pharmacology; D019913:Nuclear Localization Signals; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D016415:Sequence Alignment; D054875:Ubiquitination; D000099270:Y-Family DNA Polymerases
D004259:DNA-Directed DNA Polymerase; D009153:Mutagens; D019913:Nuclear Localization Signals; D018809:Proliferating Cell Nuclear Antigen; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases