Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Methylation-induced blocks to in vitro DNA replication.

K Larson, J Sahm, R Shenkar

    Mutation Research
    |June 1, 1985
    PubMed
    Summary
    This summary is machine-generated.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    Trajectories of health-related quality of life across age cohorts: A longitudinal analysis of the German population over 16 years.

    Social science & medicine (1982)·2025
    Same author

    Attachment styles and healthcare utilization: exploring the role of the patient-doctor relationship.

    BMC health services research·2024
    Same author

    Prevalence of pica and rumination behaviours in adults and associations with eating disorder and general psychopathology: findings form a population-based study.

    Epidemiology and psychiatric sciences·2022
    Same author

    Early weight gain in infants with Robin sequence after mandibular distraction.

    International journal of oral and maxillofacial surgery·2022
    Same author

    [Diagnostics, counselling and treatment in the context of gender incongruence, gender dysphoria and trans health : Practical information regarding the S3 guideline].

    Der Urologe. Ausg. A·2020
    Same author

    Sulfasalazine-induced mononucleosis-like-illness and haemolysis.

    Scandinavian journal of rheumatology·2020

    Methylation of DNA templates by DMS or MNNG inhibits in vitro DNA synthesis. Methylated adenine residues block DNA polymerase activity, while heating converts methylated guanine to AP sites, also halting synthesis.

    Area of Science:

    • Molecular Biology
    • Biochemistry
    • DNA Replication

    Background:

    • DNA methylation is a critical epigenetic modification.
    • Chemical modifications of DNA templates can impact DNA synthesis.
    • Understanding these impacts is crucial for molecular biology research.

    Purpose of the Study:

    • To investigate the effects of chemical methylation on DNA template function.
    • To determine how DNA polymerase activity is affected by methylated bases.
    • To elucidate the mechanism of DNA synthesis termination at modified sites.

    Main Methods:

    • In vitro DNA synthesis using M13mp2 and double-stranded DNA templates.
    • Treatment of templates with dimethyl sulfate (DMS) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG).

    Related Experiment Videos

  • Utilizing E. coli DNA polymerase I and AMV reverse transcriptase.

    • Main Results:

    • Methylation of adenine residues (A) by DMS or MNNG caused DNA synthesis termination one nucleotide downstream.
    • Heating methylated templates induced synthesis termination upstream of guanine residues (G).
    • Methylated A's were identified as direct blocks, while heating converted methylated G's to AP sites, also acting as blocks.

    Conclusions:

    • Methylated adenine residues significantly impede in vitro DNA synthesis.
    • Heating induces depurination at methylated guanine sites, creating AP sites that block DNA polymerases.
    • These findings clarify the inhibitory mechanisms of DNA methylation on DNA replication processes.