GO:1904860 DNA synthesis involved in mitotic DNA replication: Mechanism, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1904860 describes the DNA biosynthetic process that is specifically involved in mitotic DNA replication, ensuring that the genome is duplicated before mitotic cell division.
• This process is tightly coupled to S phase and is regulated by cell cycle machinery, including MYC-driven transcriptional programs.
• Key proteins include DNA polymerases, PCNA, RFC, and translesion synthesis factors such as PolDIP2 that help complete replication under stress.
• Defects in mitotic DNA replication can lead to genome instability, microcephaly, and cancer, as seen in extrachromosomal DNA maintenance and damage response coupling [1,4].
• Model organisms like Chlamydomonas reveal conserved repression of mitotic DNA replication genes during synchronous growth.
• Studying GO:1904860 requires methods such as BrdU/EdU incorporation, flow cytometry, and CRISPR-based knockout or knock-in models to dissect gene function [5,8].
Description
DNA synthesis involved in mitotic DNA replication (GO:1904860) is a biological process that encompasses any DNA biosynthetic event specifically tied to the duplication of the genome during mitotic cell division. This process is fundamental for faithful transmission of genetic information and is coordinated with the cell cycle to ensure that DNA replication occurs once and only once per cycle. Researchers study this term to understand how cells maintain genomic integrity, how replication stress is managed, and how errors contribute to diseases such as cancer and developmental disorders [1,4]. The process is not merely a housekeeping function; it is a highly regulated network that integrates signals from growth factors, DNA damage checkpoints, and developmental cues [2,8]. In this article, we dissect the molecular players, regulatory mechanisms, and experimental models used to investigate GO:1904860, with a focus on how CRISPR-based tools can accelerate discovery.
DNA synthesis involved in mitotic DNA replication At A Glance
| GO ID | GO:1904860 |
|---|---|
| GO term | DNA synthesis involved in mitotic DNA replication |
| Ontology | biological_process |
| Synonym | DNA biosynthesis involved in mitotic DNA replication; DNA formation involved in mitotic DNA replication; DNA anabolism involved in mitotic DNA replication |
| Major function | Duplication of the genome during S phase of the mitotic cell cycle |
| Related processes | DNA replication, mitotic cell cycle, S phase, DNA repair |
| Key enzymes | DNA polymerases, PCNA, RFC, translesion synthesis polymerases |
| Disease relevance | Cancer, microcephaly, genome instability disorders |
What Is GO:1904860?
GO:1904860 is defined as any DNA biosynthetic process that is involved in mitotic DNA replication. In simpler terms, it covers the synthesis of new DNA strands that occurs specifically when a cell is preparing to divide by mitosis. This includes the enzymatic steps of adding nucleotides to growing DNA chains, the proofreading and repair activities that ensure accuracy, and the coupling of these events to the mitotic cell cycle machinery. The term is a child of broader DNA replication and mitotic cell cycle processes, and it excludes DNA synthesis that occurs outside of S phase, such as repair synthesis or unscheduled replication.
Why Is DNA synthesis involved in mitotic DNA replication Important in Cell Biology?
Understanding GO:1904860 is critical because errors in mitotic DNA replication are a major source of mutations and chromosomal rearrangements that drive cancer and developmental diseases [1,4]. The process is also a target for chemotherapeutic agents like hydroxyurea, which inhibits ribonucleotide reductase and stalls replication forks. Moreover, recent evidence shows that extrachromosomal DNA replication and maintenance are coupled to DNA damage pathways in tumors, highlighting the importance of this process in cancer evolution. By studying the genes and mechanisms of GO:1904860, researchers can identify vulnerabilities for therapeutic intervention and develop biomarkers for replication stress.
• Ensures accurate duplication of the genome before mitosis, preventing aneuploidy and mutations.
• Integrates with DNA damage checkpoints to maintain genomic stability under stress [1,8].
• Dysregulation leads to cancer, as MYC-driven cell cycle control promotes replication and proliferation.
• Defects in replication genes cause congenital microcephaly and growth retardation.
• Hydroxyurea, a classic replication inhibitor, is used to treat cancers and sickle cell disease, underscoring clinical relevance.
• Endoreplication, a variant of the cell cycle, shares components with mitotic DNA replication and is important in development.
• Translesion synthesis proteins like PolDIP2 help complete replication past DNA lesions, linking GO:1904860 to DNA repair.
• Replication outside of S phase can occur in cancer and is associated with genome instability.
• Model organisms such as Chlamydomonas provide insights into the evolutionary conservation of replication control.
• CRISPR screens can identify novel regulators of mitotic DNA replication, accelerating target discovery.
What Happens During DNA synthesis involved in mitotic DNA replication?
Initiation and Licensing of Replication Origins
In simple terms: Before DNA can be copied, the cell marks starting points on the DNA and loads the copying machines.
During late mitosis and early G1, the origin recognition complex (ORC), Cdc6, and Cdt1 load the MCM2-7 helicase onto DNA to form the pre-replicative complex (pre-RC). This licensing ensures that each origin fires only once per cell cycle. The process is tightly regulated by cyclin-dependent kinases (CDKs) and is essential for mitotic DNA replication. In Chlamydomonas, CHT7 represses replication and mitotic genes during synchronous growth, highlighting conserved control mechanisms.
Activation of Replication Forks and DNA Synthesis
In simple terms: The copying machines start unwinding DNA and adding new nucleotides.
Upon entry into S phase, CDKs and DDK activate the MCM helicase, leading to unwinding of DNA and recruitment of DNA polymerases. DNA polymerase alpha primase synthesizes short RNA primers, which are extended by DNA polymerase delta and epsilon. Proliferating cell nuclear antigen (PCNA) acts as a sliding clamp, and replication factor C (RFC) loads PCNA onto DNA. This coordinated action results in leading and lagging strand synthesis [2,7].
Translesion Synthesis and Replication Stress Response
In simple terms: When the copying machine hits damage, special enzymes help it continue.
DNA lesions can stall replication forks. Translesion synthesis (TLS) polymerases, such as Pol eta, Pol iota, and Pol kappa, can bypass lesions. PolDIP2 (also known as POLDIP2) is a multifunctional protein that interacts with TLS polymerases and regulates their activity. The ATR-CHK1 pathway is activated by replication stress to stabilize forks and delay cell cycle progression, allowing time for repair [1,8].
Termination and Coupling to Mitosis
In simple terms: Once copying is done, the cell prepares to divide.
After replication forks converge, termination occurs, and the newly synthesized DNA is processed and ligated. The completion of DNA synthesis is monitored by checkpoints that ensure full replication before mitosis. Recent studies show that extrachromosomal DNA replication and maintenance are coupled with DNA damage pathways in tumors, indicating that termination and repair are intertwined. Defects in termination can lead to under-replicated regions and mitotic catastrophe.
Key Genes Involved in GO:1904860 DNA synthesis involved in mitotic DNA replication
The following genes and proteins are central to DNA synthesis involved in mitotic DNA replication, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| MCM2-7 | Helicase that unwinds DNA at replication forks | Target for replication inhibitors; marker of proliferation |
| PCNA | Sliding clamp for DNA polymerases | Therapeutic target; biomarker of replication |
| RFC | Loads PCNA onto DNA | Essential for processivity; studied in cancer |
| POLA1 | DNA polymerase alpha primase | Initiates DNA synthesis; mutations cause immunodeficiency |
| POLD1 | DNA polymerase delta | Lagging strand synthesis; proofreading; cancer predisposition |
| POLE | DNA polymerase epsilon | Leading strand synthesis; proofreading; cancer mutations |
| POLDIP2 | Multifunctional protein in translesion synthesis | Regulates TLS polymerases; potential cancer target |
| ATR | Kinase that responds to replication stress | Inhibitor in clinical trials for cancer |
| CHEK1 | Effector kinase in replication checkpoint | Target for cancer therapy |
| CDK1 | Cyclin-dependent kinase 1 | Drives mitosis; regulates replication licensing |
| CDK2 | Cyclin-dependent kinase 2 | Controls S phase entry and progression |
| MYC | Transcription factor promoting cell cycle | Overexpressed in many cancers; drives replication |
| CHT7 | Repressor of DNA replication and mitotic genes | Model for synchronous growth control |
| ORC1 | Origin recognition complex subunit | Initiates licensing; mutations in Meier-Gorlin syndrome |
| CDT1 | Licensing factor | Regulates origin firing; mutations in microcephaly |
| CDC6 | Licensing factor | Essential for pre-RC assembly |
| GMNN | Geminin, inhibitor of Cdt1 | Prevents re-replication; regulates cell cycle |
How Is DNA synthesis involved in mitotic DNA replication Regulated?
The process of DNA synthesis involved in mitotic DNA replication is regulated at multiple levels. Transcriptional control by MYC and E2F family proteins drives the expression of replication genes during G1/S transition. Post-translational modifications, such as phosphorylation by CDKs and DDK, activate replication factors. The ATR-CHK1 pathway monitors replication stress and coordinates fork stabilization with cell cycle arrest [1,8]. Additionally, the licensing system is regulated by geminin, which inhibits Cdt1 to prevent re-replication. In tumors, extrachromosomal DNA replication is coupled with DNA damage pathways, suggesting that regulation extends to non-chromosomal DNA.
DNA synthesis involved in mitotic DNA replication and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| MYC | Cancer (multiple types) | Overexpression in cell lines; knockout in cancer models |
| ORC1 | Meier-Gorlin syndrome, microcephaly | Knockout or point mutation in iPSCs |
| CDT1 | Microcephaly | Knock-in of patient mutations in neural progenitors |
| POLD1 | Colorectal cancer, polymerase proofreading-associated polyposis | Knock-in of exonuclease domain mutations |
| POLE | Colorectal cancer, endometrial cancer | Knock-in of proofreading mutations |
Cancer and Genome Instability
Dysregulated mitotic DNA replication is a hallmark of cancer. Oncogenes such as MYC promote replication stress and genomic instability, while tumor suppressors like p53 are lost, allowing cells to tolerate replication errors. Extrachromosomal DNA replication in tumors is coupled with DNA damage pathways, contributing to therapy resistance and tumor heterogeneity. Inhibitors of ATR and CHK1 are being developed to exploit replication stress in cancer cells.
Congenital Microcephaly and Developmental Disorders
Mutations in genes involved in DNA replication, such as ORC1, CDT1, and others, cause congenital microcephaly, a condition characterized by reduced brain size. These mutations impair the proliferation of neural progenitor cells, highlighting the importance of mitotic DNA replication in brain development. The study of these disorders provides insights into the tissue-specific requirements for replication factors.
Replication Stress and Chemotherapy
Hydroxyurea, a ribonucleotide reductase inhibitor, stalls DNA replication and is used to treat cancers and sickle cell disease. Understanding how cells respond to replication stress is crucial for optimizing chemotherapy and overcoming resistance. Translesion synthesis proteins like PolDIP2 are potential targets to sensitize tumors to DNA-damaging agents.
From DNA synthesis involved in mitotic DNA replication-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does gene X regulate replication origin firing? | Knockout cell lines with EdU incorporation and flow cytometry |
| Does mutation Y affect replication fork speed? | Point mutation knock-in followed by DNA fiber assays |
| Does protein Z localize to replication forks? | Tagged knock-in with GFP or HA for imaging |
| Does overexpression of oncogene W cause replication stress? | Overexpression cell lines with ATR/CHK1 activation markers |
| Is gene V essential for S phase progression? | CRISPR knockout with cell cycle analysis |
| Does gene U interact with PCNA? | Knock-in of affinity tags for proteomics |
How to Study the DNA synthesis involved in mitotic DNA replication Process
| Method | What It Measures | Typical Application |
|---|---|---|
| EdU/BrdU incorporation | DNA synthesis rate and S phase fraction | Assessing replication defects in knockout cells |
| DNA fiber assay | Fork speed, stalling, and restart | Studying replication stress responses |
| Flow cytometry | Cell cycle distribution | Evaluating G1/S transition and mitosis |
| CRISPR knockout screen | Genes required for replication or drug resistance | Identifying novel regulators |
| Affinity proteomics | Protein-protein interactions | Mapping replication complexes |
| Immunofluorescence | Localization of replication proteins | Visualizing foci formation |
| Western blot | Protein expression and phosphorylation | Checking checkpoint activation |
| qPCR | DNA copy number and replication intermediates | Measuring replication efficiency |
Measuring DNA Synthesis with Nucleotide Analogs
BrdU or EdU incorporation followed by flow cytometry or microscopy is a standard method to measure DNA synthesis in S phase. These assays can quantify the fraction of cells replicating and detect perturbations in mitotic DNA replication. Combined with cell cycle inhibitors, they allow dissection of specific stages.
DNA Fiber Assays for Fork Dynamics
DNA fiber assays visualize individual replication forks by labeling newly synthesized DNA with sequential halogenated nucleotides. This technique measures fork speed, stalling, and restart, providing insights into replication stress and the role of specific genes.
CRISPR Screens for Replication Regulators
Genome-wide CRISPR knockout or activation screens can identify genes that are essential for mitotic DNA replication or that confer resistance to replication inhibitors. Such screens have uncovered novel factors in DNA damage response and replication.
Proteomics and Interactomics
Affinity purification of replication proteins followed by mass spectrometry reveals dynamic interactomes. For example, tagging PCNA or PolDIP2 can identify partners involved in translesion synthesis. These methods help build a comprehensive map of the replication machinery.
How CRISPR Can Be Used to Study GO:1904860 DNA synthesis involved in mitotic DNA replication
Knockout
CRISPR knockout of genes involved in mitotic DNA replication can reveal essential functions. For example, knocking out MCM subunits or PCNA leads to replication failure and cell cycle arrest. Knockout models are valuable for identifying synthetic lethal interactions with DNA damage response genes.
Point Mutation
Introducing specific point mutations, such as in the exonuclease domain of POLD1 or POLE, mimics cancer-associated mutations and allows study of proofreading defects. Point mutation knock-in can also model microcephaly-associated mutations in ORC1 or CDT1.
Knock-in
Knock-in of tagged versions of replication proteins, such as GFP-PCNA or HA-PolDIP2, enables live-cell imaging and proteomic analysis. Knock-in of patient-derived mutations into endogenous loci provides physiologically relevant disease models.
Overexpression
Overexpression of oncogenes like MYC or replication factors can induce replication stress and genomic instability, modeling early cancer events. Overexpression studies help determine sufficiency of a gene to drive replication phenotypes.
How EDITGENE Supports DNA synthesis involved in mitotic DNA replication Research
Researchers studying DNA synthesis involved in mitotic DNA replication-related genes often need to determine whether a candidate gene is causally involved in replication control, fork stability, or disease progression. EDITGENE provides a comprehensive suite of CRISPR services to generate precisely engineered cell models, enabling functional validation and mechanistic studies.
Contact EDITGENE today to design your custom CRISPR model for DNA synthesis involved in mitotic DNA replication research.
Frequently Asked Questions About DNA synthesis involved in mitotic DNA replication
What is GO:1904860?
GO:1904860 is a Gene Ontology term for any DNA biosynthetic process that is involved in mitotic DNA replication, ensuring genome duplication before cell division.
What genes are involved in DNA synthesis involved in mitotic DNA replication?
Key genes include MCM2-7, PCNA, RFC, POLA1, POLD1, POLE, POLDIP2, ATR, CHEK1, CDK1, CDK2, MYC, ORC1, CDT1, CDC6, and GMNN [2,7].
How is mitotic DNA replication regulated?
It is regulated by CDKs, DDK, MYC, E2F, and checkpoint kinases like ATR and CHK1, which coordinate origin firing and stress responses [1,2].
What diseases are associated with defects in mitotic DNA replication?
Cancer, congenital microcephaly, and genome instability disorders are linked to replication defects [1,4].
What methods are used to study DNA synthesis in mitosis?
EdU/BrdU incorporation, DNA fiber assays, flow cytometry, CRISPR screens, and proteomics are commonly used [5,8].
How can CRISPR help study GO:1904860?
CRISPR knockout, knock-in, point mutation, and overexpression models allow functional dissection of genes involved in replication [2,7].
What is the role of PolDIP2 in DNA replication?
PolDIP2 is a multifunctional protein that regulates translesion synthesis polymerases, helping replication bypass DNA lesions.
Can replication occur outside of S phase?
Yes, recent studies show that genome replication can be completed outside of S phase, which is relevant to cancer and genome instability.
What is the connection between MYC and DNA replication?
MYC promotes cell cycle progression and drives expression of replication genes, contributing to replication stress in cancer.
How does hydroxyurea affect mitotic DNA replication?
Hydroxyurea inhibits ribonucleotide reductase, depleting nucleotides and stalling replication forks, leading to S phase arrest.
Conclusion
GO:1904860 DNA synthesis involved in mitotic DNA replication is a fundamental biological process that ensures accurate genome duplication. Its dysregulation is implicated in cancer, microcephaly, and other diseases, making it a rich area for research. By leveraging CRISPR-based models and advanced screening methods, researchers can uncover new regulators and therapeutic targets. EDITGENE provides the tools and expertise to accelerate these discoveries.
References
- 1. Kang X et al.. 2025. Extrachromosomal DNA replication and maintenance couple with DNA damage pathway in tumors.. Cell 188(13):3405-3421.e27 PMID: 40300601
- 2. Bretones G et al.. 2015. Myc and cell cycle control.. Biochim Biophys Acta 1849(5):506-16 PMID: 24704206
- 3. Lin YT et al.. 2022. Chlamydomonas CHT7 is involved in repressing DNA replication and mitotic genes during synchronous growth.. G3 (Bethesda) 12(3) PMID: 35137070
- 4. Alcantara D et al.. 2014. Congenital microcephaly.. Am J Med Genet C Semin Med Genet 166C(2):124-39 PMID: 24816482
- 5. Timson J. 1975. Hydroxyurea.. Mutat Res 32(2):115-32 PMID: 765790
- 6. Zielke N et al.. 2013. Endoreplication.. Cold Spring Harb Perspect Biol 5(1):a012948 PMID: 23284048
- 7. Gagarinskaya DI et al.. 2020. A Multifunctional Protein PolDIP2 in DNA Translesion Synthesis.. Adv Exp Med Biol 1241:35-45 PMID: 32383114
- 8. Bhowmick R et al.. 2023. Completing genome replication outside of S phase.. Mol Cell 83(20):3596-3607 PMID: 37716351
