Coordinated cell cycle progression is a complex challenge for eukaryotic cells. In response to DNA damage, a plethora of molecular signals must be integrated to establish cell cycle checkpoints, allowing time for DNA repair.
Phospho-Ser/Thr-binding domains act as central regulators (hubs) that integrate the activity of multiple kinase signalling cascades to coordinate cell cycle progression and checkpoint control following genotoxic stress.
Phospho-Ser/Thr-binding domains, such as 14-3-3 proteins, WW domains, Polo-box domains, WD40 repeats, BRCA1 carboxy-terminal (BRCT) domains and forkhead-associated (FHA) domains can be found in numerous proteins involved in cell cycle regulation and the DNA damage response and are conserved from metazoans to the simplest single cell eukaryotes.
14-3-3 proteins control mitotic progression and the G2–M checkpoint by regulating the subcellular localization and the catalytic activity of their phosphorylated client proteins.
The WW domain-containing protein PIN1 slows progression through mitosis and is also required for mitotic exit and the intra-S phase checkpoint.
Polo-box domains are a hallmark feature of Polo-like kinases (PLKs) and help localize PLKs to their substrates and regulate the activity of their kinase domains. PLK1 activity is required for the timely activation of CDC25B and CDC25C to initiate mitosis, for the coordinated progression of cells through mitosis and for inactivating DNA damage checkpoints once repair is complete.
WD40 domains can be found in a subset of F-box proteins, including βTrCP (β-transducin repeat-containing protein), CDC4 and FBW7.
SCFβTrCP (SKP1–cullin 1–F-box complex containing βTrCP)-dependent substrate degradation controls two opposing cellular responses following genotoxic stress: checkpoint-activating degradation of CDC25A following DNA damage and checkpoint-inactivating claspin and WEE1 degradation following damage repair.
BRCT domains are present in various DNA replication and repair proteins, including BRCA1, MDC1 (mediator of DNA damage checkpoint protein 1), 53BP1 (p53 binding protein 1), TOPBP1 (topoisomerase-binding protein 1), PTIP (PAX-interacting protein 1), XRCC1 (X-ray repair cross-complementing 1) and DNA ligase IV. BRCT domain-containing proteins control numerous diverse processes, including homologous recombination-mediated DNA double strand repair and base excision repair, as well as cell cycle checkpoint activation and maintenance.
FHA domains are found in proteins involved in transcription, DNA damage repair and cell cycle control, as well as in kinesin-like motors and regulators of small G proteins. Within the DNA damage response cascade, FHA domain-containing proteins, such as RING finger 8 (RNF8) and MDC1 control the assembly of signalling complexes at the site of the DNA lesion, whereas the FHA domain-containing checkpoint kinase CHK2 acts as a mobile messenger within the nucleoplasm.
Coordinated progression through the cell cycle is a complex challenge for eukaryotic cells. Following genotoxic stress, diverse molecular signals must be integrated to establish checkpoints specific for each cell cycle stage, allowing time for various types of DNA repair. Phospho-Ser/Thr-binding domains have emerged as crucial regulators of cell cycle progression and DNA damage signalling. Such domains include 14-3-3 proteins, WW domains, Polo-box domains (in PLK1), WD40 repeats (including those in the E3 ligase SCFβTrCP), BRCT domains (including those in BRCA1) and FHA domains (such as in CHK2 and MDC1). Progress has been made in our understanding of the motif (or motifs) that these phospho-Ser/Thr-binding domains connect with on their targets and how these interactions influence the cell cycle and DNA damage response.
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The authors apologize to their colleagues for the omission of many seminal contributions to the field, and their corresponding references, owing to space constraints. They gratefully acknowledge S. J. Smerdon, their long-standing collaborator, for his continual insights and encouragement in these projects. This work was supported by the US National Institutes of Health (GM68762, CA112967, ES015339, and ES020466 to M.B.Y.), the Volkswagenstiftung (Lichtenberg Program; to H.C.R.), the Deutsche Forschungsgemeinschaft (KFO-286, SFB-832, SFB-829, RE2246/2-1; to H.C.R.), the Helmholtz-Gemeinschaft (Preclinical Comprehensive Cancer Center; to H.C.R.), the Deutsche Jose Carreras Stiftung (DJCLS-R12/26 to H.C.R) and the David H. Koch Fund (H.C.R. and M.B.Y.).
The authors declare no competing financial interests.
- Cap-dependent mRNA translation
Describes a process by which mRNA translation is initiated through the recruitment of 40S ribosomes to the 5′ 7-methylguanosine-capped end of mRNAs via eukaryotic translation initiation factor 4F (eIF4F).
- Cap-independent mRNA translation
Describes a process of mRNA translation by which the requirement for the 5′ 7-methylguanosine cap and eukaryotic translation initiation factor 4E (eIF4E) is bypassed, and translation initiation occurs instead through ribosome recruitment to internal ribosomal entry sites (IRESs).
A dense protein structure that represents the remnant of the spindle midzone located at the centre of the narrow membranous bridge connecting two dividing cells at the end of cytokinesis.
A protein complex consisting of MKLP1 (mitotic kinesin-like protein 1) and CYK4 (cytokinesis defect 4) that controls mitosis through various mechanisms, including initiation of central spindle assembly, positioning of the division plane and regulation of midbody assembly and abscission.
- Spindle midzone
A region of the anaphase spindle that is composed of overlapping antiparallel microtubules from opposite spindle poles. It is also known as the central spindle.
- Intra-S phase checkpoint
ATM (ataxia-telangiectasia mutated)- and ATR-dependent, transient inhibition of DNA replication in response to DNA damage. Defects in an ionizing radiation-induced intra-S phase checkpoint cause radioresistant DNA synthesis.
- K m
(Michaelis constant). An intrinsic property of enzymes that describes the concentration for a particular substrate at which the enzyme works at half-maximal capacity, that is, at Vmax/2.
- Spindle checkpoint
A mechanism that detects unattached kinetochores in mitosis and arrests the cell cycle in metaphase, prior to anaphase onset.
- E3 ubiquitin ligases
These ligases facilitate the transfer of ubiquitin from a ubiquitin conjugating enzyme (E2) to Lys residues on a substrate, often forming polyubiquitin chains that target the substrate for degradation by the proteasome.
- Homologous recombination-mediated DSB repair
A DNA recombination pathway that includes the repair of DNA double-strand breaks (DSBs). This pathway involves the resection of one strand of the DNA in the vicinity of the break followed by use of a homologous double-stranded DNA molecule as a template for the repair of the broken DNA.
- Nucleotide excision repair
This repair pathway is used to remove the vast majority of lesions located on a DNA single strand, including lesions caused by ultraviolet (UV) light and cisplatin damage. This pathway involves the enzymatic excision of one or more nucleotides from the damaged DNA single strand followed by new DNA synthesis using the opposing strand as a template.
- Non-homologous end-joining
(NHEJ). The main DNA repair pathway used throughout the cell cycle to repair chromosomal double-strand DNA breaks in somatic cells, involving the fusion of the two broken DNA ends. In contrast to homologous recombination-mediated repair, NHEJ is error-prone because it leads to the joining of heterologous ends, often with the loss of intervening genetic material in the region of the break.
- Base excision repair
(BER). A DNA repair pathway in which a damaged DNA base within a single DNA strand is initially removed by a glycosylase enzyme and the abasic nucleotide is then replaced, along with a short patch of adjacent nucleotides.
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Reinhardt, H., Yaffe, M. Phospho-Ser/Thr-binding domains: navigating the cell cycle and DNA damage response. Nat Rev Mol Cell Biol 14, 563–580 (2013). https://doi.org/10.1038/nrm3640
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