# THE BIOCHEMISTRY OF DNA ALKYL PHOSPHOTRIESTERS

> **NIH NIH R01** · TEMPLE UNIV OF THE COMMONWEALTH · 1988 · $65,229

## Abstract

Quantitatively the greatest DNA oxygen lesion produced by a family of
1-alkyl-1-nitroso compound carcinogens, which includes dimethyl and
diethylnitrosamine, methyl and ethylnitrosourea and
1-methyl-2-nitro-1-nitrosoguanidine, is at the backbone phosphate groups.
The yield is 2-to 5-fold higher than at the 06-position of guanine.  DNA
oxygen alkylation correlates with carcinogenic potential.  It is not known
to what degree these alkyl phosphotriesters effect the biochemistry of DNA;
I am considering the possibility that they aggravate the binding of
proteins which function to insure the proper transmission of genetic
information.  Using an automatic DNA synthesizer and the Beta-cyanoethyl
phosphoramidite chemistry I propose to synthesize DNA polymers of known
length and sequence containing any percentage of diastereomerically pure
ethyl phosphotriesters.  With the view that phosphotriesters could perturb
the DNA conformational signals which define protein recognition sites, I
will consider the effects of these lesions on DNA helical conformation and
conformational stability using helix-coil transition experiments and
circular dichroism.  I will assess the effects of phosphotriesters on the
activity of the single strand binding protein, T4 phage gene 32 protein.
Perturbations in the intrinsic binding constant and in the cooperativity
parameter will be evaluated by taking advantage of tryptophan fluorescence
quenching.  The efficiency of E. coli polymerase I on
phosphotriester-containing templates will be tested.  I will examine the
polymerization rate, nucleoside triphosphate turnover and nucleotide
incorporation fidelity utilizing standard protocols and HPLC analysis.
Finally I will study the effects of phosphotriesters placed at known
contact positions of the lac UV5 promoter on the efficiency of E. coli RNA
polymerase message initiation using the abortive initiation assay.  The
promoter fragment will be either synthesized de novo or produced by
utilizing some of the methodology developed for oligonucleotide-directed
site-specific mutagenesis.

## Key facts

- **NIH application ID:** 3176097
- **Project number:** 5R01CA038077-04
- **Recipient organization:** TEMPLE UNIV OF THE COMMONWEALTH
- **SAM.gov UEI:** QD4MGHFDJKU1
- **Principal Investigator:** DAVID E JENSEN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $65,229
- **Award type:** 5
- **Project period:** 1984-07-01 → 1990-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3176097

## Citation

> US National Institutes of Health, RePORTER application 3176097, THE BIOCHEMISTRY OF DNA ALKYL PHOSPHOTRIESTERS (5R01CA038077-04). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3176097. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*