Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Transferase other than ribonuclease
Patent
1998-01-20
2000-10-24
Patterson, Jr., Charles L.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Transferase other than ribonuclease
C12N 912
Patent
active
061365823
ABSTRACT:
This invention provides a polypeptide capable of polymerizing i) deoxyribonucleotides; ii) ribonucleotides; or iii) one or more deoxyribonucleotides and one or more ribonucleotides. This invention further provides a purified Moloney murine leukemia virus reverse transcriptase, wherein an amino acid corresponding to position 155 of a wild type Moloney murine leukemia virus reverse transcriptase is a valine. This invention also provides a method of polymerizing a nucleic acid molecule comprising one or more deoxyribonucleotides and one or more ribonucleotides comprising: contacting a polypeptide capable of polymerizing i) deoxyribonucleotides; ii) ribonucleotides; or iii) one or more deoxyribonucleotides and one or more ribonucleotides with deoxyribonucleotides and ribonucleotides under conditions permitting incorporation of deoxyribonucleotides and ribonucleotides into a nucleic acid molecule. This invention still further provides a method of converting a DNA polymerase into an RNA polymerase comprising: a) structurally modifying the DNA polymerase; b) contacting the structurally modified DNA polymerase of step (a) with deoxyribonucleotides and ribonucleotides under conditions permitting incorporation of deoxyribonucleotides and ribonucleotides into a nucleic acid molecule; and c) detecting polymerization of ribonucleotides in the nucleic acid molecule resulting from step (b), the detection of ribonucleotides in the polymerized nucleic acid molecule indicating that the structurally modified DNA polymerase has been converted into an RNA polymerase. This invention also provides a nucleic acid molecule polymerized by the above-described method of polymerizing a nucleic acid molecule comprising one or more deoxyribonucleotides and one or more ribonucleotides.
REFERENCES:
Arnold, E. et al., (1995) "Structures of DNA and RNA Polymerases and Their Interactions With Nucleic Acid Substrates," Current Opinion In Structural Biology, 5(1):27-38.(Exhibit 1).
Baltimore, D., (1970) "Viral RNA-dependent DNA Polymerase," Nature, 226(5252): 1209-1211 (Exhibit 2).
Delarue, M. et al., (1990) "An Attempt to Unify the Structure of Polymerases," Protein Eng., 3(6):461-467. (Exhibit 3).
Georgiadis, M.M. et al., (1995) "Mechanistic Implications From the Structure of a Catalytic Fragment of Moloney Murine Leukemia Virus Reverse Transcriptase," Structure, (London), 3(9):879-892. (Exhibit 4).
Goff, S. et al., (1981) "Isolation and Properties of Moloney Murine Leukemia Virus Mutants: Use of a Rapid Assay for Release of Virion Reverse Transcriptase," J. Virology, 38(1):239-248. (Exhibit 5).
Joyce, C.M. and Steitz, T.A.,-(1994) "Function and Structure Relationships in DNA Polymerases," Annu. Rev. Biochem., 63:777-822. (Exhibit 6).
Joyce, C.M., (1997) "Choosing the Right Sugar: How Polymerases Select a Nucleotide Substrate," Proc. Natl. Acad. Sci. USA, 94(5):1619-1622. (Exhibit 7).
Patel, P.H. et al., (1995) "Insights Into DNA Polymerization Mechanisms From Structure and Function Analysis of HIV-1 Reverse Transcriptase," Biochemistry, 34(16):5351-5363. (Exhibit 8).
Sousa, R. et al., (1993) "Crystal Structure of Bacteriophage T7 RNA Polymerase at 3.3 .ANG. Resolution," Nature, 364(6438):593-599. (Exhibit 9).
Sousa, R. and Padilla, R., (1995) "A Mutant T7 RNA Polymerase as a DNA Polyerase," EMBO J., 14(18):4609-4621. (Exhibit 10).
Steitz, T.A. et al., (1994) "A Unified Polymerase Mechanism for Nonhomologous DNA and RNA Polymerases," Science, 266(5913):2022-2025. (Exhibit 11).
Temin, H. and Mizutani, S., (1970) "RNA-dependent DNA Polymerase in Virions of Rous Sarcoma Virus," Nature, (London), 226(5252):1211-1213. (Exhibit 12).
Gao, Guangxia, et al. (1997) Proc Natl. Acad, Sci, USA 94, 407-411.
Gao Guangxia
Goff Stephen P.
Patterson Jr. Charles L.
The Trustees of Columbia University in the City
White John P.
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