# Initiation of DNA Replication at Cell Origins in Yeast

> **NIH NIH R01** · COLD SPRING HARBOR LABORATORY · 2020 · $729,600

## Abstract

The goal of this research is to determine the mechanism and regulation of the initiation of DNA
replication in eukaryotic cells. It is clear that for maintenance of the integrity of the genome from
one cell generation to the next, DNA and its associated chromatin structures must be duplicated
in a highly controlled and accurate manner. Interruption of these controls may promote genome
instability and lead to neoplastic transformation in somatic cells or result in mutations in the germ
line that can cause many different disorders. Moreover, the DNA replication proteins represent
tangible targets for therapeutic intervention and diagnosis of proliferation of cancer cells, and
other proliferative disorders. The initiator protein (ORC) cooperates with a series of DNA
replication proteins, including Cdc6, Cdt1 and the MCM2-7 hexamer to establish at origins of DNA
replication a pre-Replicative Complex (pre-RC) that facilitates later initiation of DNA synthesis at
each origin. Recent progress has enabled the assembly of the pre-RC in vitro with purified
proteins. The proposed research in this application will investigate, using the yeast S. cerevisiae
and human cell proteins, how the initiation of DNA replication occurs following pre-RC assembly
and how this process is regulated by the Cdc7-Dbf4 (DDK) protein kinase and by an intrinsic
inhibitor of initiation of DNA replication within the Mcm4 subunit of the MCM2-7 complex.
Biochemical and structural studies will elucidate how molecular machines that are driven by ATP
function to load ring shaped protein onto double stranded DNA and how origin recognition has
evolved in eukaryotes.

## Key facts

- **NIH application ID:** 9858362
- **Project number:** 5R01GM045436-28
- **Recipient organization:** COLD SPRING HARBOR LABORATORY
- **Principal Investigator:** BRUCE W. STILLMAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $729,600
- **Award type:** 5
- **Project period:** 1991-07-01 → 2022-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9858362

## Citation

> US National Institutes of Health, RePORTER application 9858362, Initiation of DNA Replication at Cell Origins in Yeast (5R01GM045436-28). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9858362. Licensed CC0.

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