Excision of b - D - and b - L -Nucleotide Analogs from DNA by the Human Cytosolic 3 9 -to-5 9 Exonuclease

The cytosolic 3 9 -to-5 9 exonuclease from chronic lymphocytic leukemia cells was highly purified, and its ability to remove b - D and b - L -nucleotide analogs from the 3 9 -end of DNA was deter-mined. The relative rate of excision of b - D -ddCMP, b - L -ddCMP, b - L -FddCMP, b - L -SddCMP, b - L -Fd4CMP, and b - L -OddCMP from the 3 9 -end of a single-stranded oligonucleotide primer or a primer annealed with complementary DNA and/or RNA tem- plates was assessed. The rate of excision of b - D -nucleotides from the 3 9 -end of DNA was higher than that of b - L -nucleotides, which could be partly attributable to the affinity of the enzyme to b - D -nucleotide-terminated DNA being 5-fold higher compared with that of b - L -nucleotide-terminated DNA. The rate of removal of b - L -Fd4CMP and b - L -OddCMP from the 3 9 -end of DNA was at least 8 to 10 times lower compared with that of b - L -SddCMP. HIV reverse transcriptase could elongate DNA primers after the removal of chain terminators by the cytosolic exonuclease. Concentrations of nucleoside 5 9 -monophosphate analogs that inhibit the cytosolic exonuclease by 50% were estimated. Among the nucleoside 5 9 -monophosphate analogs examined, b - L -Fd4CMP appeared to be the most effective inhibitor of

[1]  A. So,et al.  Unblocking of chain-terminated primer by HIV-1 reverse transcriptase through a nucleotide-dependent mechanism. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Shwu-Huey Liu,et al.  Anti-Hepatitis B Virus Activity and Metabolism of 2′,3′-Dideoxy-2′,3′-Didehydro-β-l(−)-5-Fluorocytidine , 1998, Antimicrobial Agents and Chemotherapy.

[3]  M. Kukhanova,et al.  Metabolism of 2′,3′-Dideoxy-2′,3′-Didehydro-β-l(−)-5-Fluorocytidine and Its Activity in Combination with Clinically Approved Anti-Human Immunodeficiency Virus β-d(+) Nucleoside Analogs In Vitro , 1998, Antimicrobial Agents and Chemotherapy.

[4]  E. Gullen,et al.  Design and Synthesis of 2′,3′‐Dideoxy‐2′,3′‐didehydro‐β‐L‐ cytidine (β‐L‐d4C) and 2′,3′‐Dideoxy‐2′,3′‐didehydro‐β‐L‐5‐ fluorocytidine (β‐L‐Fd4C), Two Exceptionally Potent Inhibitors of Human Hepatitis B Virus (HBV) and Potent Inhibitors of Human Immunodeficiency Virus (HIV) in vitro. , 1996 .

[5]  M. Chun,et al.  Dioxolane cytosine nucleosides as anti-hepatitis B agents , 1995 .

[6]  V. Skalski,et al.  Removal of anti-human immunodeficiency virus 2',3'-dideoxynucleoside monophosphates from DNA by a novel human cytosolic 3'-->5' exonuclease. , 1995, Biochemical pharmacology.

[7]  G. Dutschman,et al.  Studies of the Pharmacokinetics and Toxicology of 2′,3′-Dideoxy-β-L-5-fluorocytidine (β-L-FddC) and 2′,3′-Dideoxy-β-L-cytidine (β-L-ddC) In Vivo; and Synthesis and Antiviral Evaluations of 2′,3′-Dideoxy-β-L-5-azacytidine , 1995 .

[8]  Y. Cheng,et al.  Synthesis and biological evaluation of 2',3'-dideoxy-L-pyrimidine nucleosides as potential antiviral agents against human immunodeficiency virus (HIV) and hepatitis B virus (HBV). , 1994, Journal of medicinal chemistry.

[9]  G. Dutschman,et al.  Antiviral activity of 2′,3′-dideoxy-β-L-5-fluorocytidine (β-L-FddC) and 2′,3′-dideoxy-β-L-cytidine (β-L-ddC) against hepatitis B virus and human immunodeficiency virus type 1 in vitro , 1994 .

[10]  V. Skalski,et al.  The biochemical basis for the differential anti-human immunodeficiency virus activity of two cis enantiomers of 2',3'-dideoxy-3'-thiacytidine. , 1993, The Journal of biological chemistry.

[11]  I. Shevelev,et al.  Proof-reading 3'-->5' exonucleases isolated from rat liver nuclei. , 1993, European journal of biochemistry.

[12]  V. Skalski,et al.  Biochemical pharmacology of (+)- and (-)-2',3'-dideoxy-3'-thiacytidine as anti-hepatitis B virus agents. , 1992, The Journal of biological chemistry.

[13]  R. Schinazi,et al.  Deoxycytidine deaminase-resistant stereoisomer is the active form of (+/-)-2',3'-dideoxy-3'-thiacytidine in the inhibition of hepatitis B virus replication. , 1992, The Journal of biological chemistry.

[14]  R. Schinazi,et al.  Inhibition of the replication of hepatitis B virus in vitro by 2',3'-dideoxy-3'-thiacytidine and related analogues. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[15]  M. Goodman,et al.  Base mispair extension kinetics. Comparison of DNA polymerase alpha and reverse transcriptase. , 1990, The Journal of biological chemistry.

[16]  W. Skarnes,et al.  Exonuclease activity associated with a multiprotein form of HeLa cell DNA polymerase alpha. Purification and properties of the exonuclease. , 1986, The Journal of biological chemistry.

[17]  D W Barry,et al.  3'-Azido-3'-deoxythymidine (BW A509U): an antiviral agent that inhibits the infectivity and cytopathic effect of human T-lymphotropic virus type III/lymphadenopathy-associated virus in vitro. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[18]  C. Kwan.  A cytoplasmic exoribonuclease from HeLa cells. , 1977, Biochimica et biophysica acta.