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Transcription factor OTF-1 is functionally identical to the DNA replication factor NF-III
E A O'Neill1, C Fletcher, C R Burrow
1Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Science (New York, N.Y.)
|September 2, 1988
Summary
Octamer transcription factor-1 (OTF-1) and nuclear factor III (NF-III) are the same protein. This single protein, NF-III/OTF-1, plays a crucial role in both cellular DNA replication and transcription.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Octamer transcription factor-1 (OTF-1) and nuclear factor III (NF-III) are known sequence-specific DNA binding proteins.
- OTF-1 is involved in transcription activation, while NF-III is involved in DNA replication activation.
Purpose of the Study:
- To investigate the relationship between OTF-1 and NF-III.
- To determine if OTF-1 and NF-III are distinct proteins or the same entity.
Main Methods:
- SDS-polyacrylamide gel electrophoresis to compare protein mobility.
- DNA binding assays to assess binding site and affinity to the adenovirus origin of replication.
- In vitro assays to test the functional substitution of OTF-1 for NF-III in DNA replication and vice versa for transcription.
Main Results:
- OTF-1 and NF-III exhibit identical mobilities on SDS-PAGE.
- OTF-1 binds to the adenovirus origin of replication at the same site and affinity as NF-III.
- OTF-1 can functionally replace NF-III in initiating viral DNA replication, and NF-III can replace OTF-1 in activating histone H2b transcription.
Conclusions:
- OTF-1 and NF-III are physically and biologically indistinguishable.
- The protein, identified as NF-III/OTF-1, functions in both cellular DNA replication and transcription activation.
- This finding suggests a dual role for a single protein in fundamental cellular processes.