# EXCISION REPAIR OF UV IRRADIATED DNA IN YEAST

> **NIH NIH R01** · UNIVERSITY OF ROCHESTER · 1987 · $136,446

## Abstract

Our overall objective is to clone the RAD1, RAD2, RAD3 and RAD4 gene of the
yeast, Saccharomyces cerevisiae, which function in incision of DNA
containing ultraviolet light (UV) induced pyrimidine dimers and to study
their structure, regulation of expression and protein products.  The RAD
genes are being cloned either by complementation in yeast or by overlap
hybridization.  We will verify the identity of the DNA fragments
complementing the rad mutants by integrating and mapping them to their
chromosomal site in the yeast genome.  Fine structure restriction maps of
RAD genes will be obtained and each gene subcloned to contain the smallest
flanking sequence.  We will determine the mRNA sizes of the RAD genes by
using RNA gel blots and enquire if there is induction of RAD genes at the
transcriptional level following UV irradiation or treatment with other DNA
damaging agents.  The direction of transcription of the RAD genes will be
determined by using the RAD DNA fragments cloned in phage M13 as probes.
The 5 foot termini of in vivo RAD transcripts will be located by S1
nuclease mapping.  The nucleotide sequence of the RAD genes will be
determined by the Sanger dideoxy method.  We will check if we can identify
the RAD proteins in the E. coli maxicell system or in yeast cells
containing the RAD genes on high copy number plasmids.  Fusions of RAD
genes with the E. coli lacz gene will be constructed to study regulation at
the translational level and to purify the RAD proteins.  The purified RAD
proteins will be characterized for DNA binding, ATPase and UV endonuclease
activity.  In humans, xeroderma pigmentosum patients are defective in
removing UV induced pyrimidine dimers from DNA and show an increased
frequency of cancers.  Other human genetic diseases are also associated
with defective DNA repair and increased carcinogenesis.  Therefore, it is
important to understand the molecular mechanisms of DNA repair in
eukaryotes.  The proposed work should provide a model system for
understanding the molecular and genetic mechanisms of incision of UV
irradiated DNA in eukaryotes, including humans.

## Key facts

- **NIH application ID:** 3172787
- **Project number:** 5R01CA035035-05
- **Recipient organization:** UNIVERSITY OF ROCHESTER
- **SAM.gov UEI:** F27KDXZMF9Y8
- **Principal Investigator:** SATYA PRAKASH
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $136,446
- **Award type:** 5
- **Project period:** 1983-07-01 → 1988-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3172787

## Citation

> US National Institutes of Health, RePORTER application 3172787, EXCISION REPAIR OF UV IRRADIATED DNA IN YEAST (5R01CA035035-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3172787. Licensed CC0.

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