Boranophosphate backbone: a mimic of phosphodiesters, phosphorothioates, and methyl phosphonates

Shaw, Barbara Ramsay; Sergueev, Dmitri; He, Kebo; Porter, Ken; Summers, Jack S.; Sergueeva, Zinaida A.; Rait, Vladimir

doi:10.1016/s0076-6879(00)13015-0

Cited by 55 publications

(31 citation statements)

References 48 publications

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“…Acyclovir-TP and d4TTP were obtained from Sierra Bioresearch (Tucson, AZ). The ␣-boranophosphate derivative of d4TTP (␣-borano-d4TTP) was synthesized as described previously (38,39), and its R p and S p diastereoisomers were purified by reverse-phase high pressure liquid chromatography (40). Before use, nucleoside TPs were treated with inorganic pyrophosphatase (Roche Applied Science) to remove traces of PP i , as described (41).…”

Section: Methodsmentioning

confidence: 99%

Thymidine Analogue Resistance Suppression by V75I of HIV-1 Reverse Transcriptase

Matamoros

Nevot

Martı́nez

et al. 2009

Journal of Biological Chemistry

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Val75 of HIV-1 reverse transcriptase (RT) plays a role in positioning the template nucleotide ؉1 during the formation of the ternary complex. Mutations, such as V75M and V75A, emerge in patients infected with HIV-1 group M subtype B and group O variants, after failing treatment with stavudine (d4T) and other nucleoside RT inhibitors. V75I is an accessory mutation of the Q151M multidrug resistance complex of HIV-1 RT and is rarely associated with thymidine analogue resistance mutations (TAMs). In vitro, it confers resistance to acyclovir. TAMs confer resistance to zidovudine (AZT) and d4T by increasing the rate of ATP-mediated excision of the terminal nucleotide monophosphate (primer unblocking). In a wild-type HIV-1 group O RT sequence context, V75A and V75M conferred increased excision activity on d4T-terminated primers, in the presence of PP i . In contrast, V75I decreased the PP i -mediated unblocking efficiency on AZT and d4T-terminated primers, in different sequence contexts (i.e. wild-type group M subtype B or group O RTs). Interestingly, in the sequence context of an excision-proficient RT (i.e. M41L/A62V/T69SSS/K70R/ T215Y), the introduction of V75I led to a significant decrease of its ATP-dependent excision activity on AZT-, d4T-, and acyclovir-terminated primers. The excision rate of d4T-monophosphate in the presence of ATP (3.2 mM) was about 10 times higher for M41L/A62V/T69SSS/K70R/T215Y than for the mutant M41L/A62V/T69SSS/K70R/V75I/T215Y RT. The antagonistic effect of V75I with TAMs was further demonstrated in phenotypic assays. Recombinant HIV-1 containing the M41L/A62V/ T69SSS/K70R/V75I/T215Y RT showed 18.3-and 1.5-fold increased susceptibility to AZT and d4T, respectively, in comparison with virus containing the M41L/A62V/T69SSS/K70R/ T215Y RT.

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Section: Methodsmentioning

confidence: 99%

Thymidine Analogue Resistance Suppression by V75I of HIV-1 Reverse Transcriptase

Matamoros

Nevot

Martı́nez

et al. 2009

Journal of Biological Chemistry

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“…AZTTP and d4TTP were purchased from Moravek Biochemicals and Sierra Research, respectively. a-BH 3 -AZTTP and a-BH 3 -d4TTP were synthesized as described, 48,49 and their R p and S p diastereoisomers were purified by reversed-phase HPLC. 26 AZTTPaS was obtained from TriLink BioTechnologies (San Diego, CA) and its S p and R p diastereoisomers were separated by reversed-phase HPLC on a Vydac C 18 218TP54 column under isocratic elution conditions (5%(v/v) acetonitrile in 10 mM triethylammonium acetate buffer (pH 6.8)).…”

Section: Nucleotides and Template-primersmentioning

confidence: 99%

Suppression of Multidrug-resistant HIV-1 Reverse Transcriptase Primer Unblocking Activity by α-Phosphate-modified Thymidine Analogues

Matamoros

Deval

Guerreiro

et al. 2005

Journal of Molecular Biology

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“…Such oligonucleotides are resistant to exonucleases and may find applications in direct PCR product sequencing (8 -11). Oligonucleotides bearing a borano (BH 3 ) group on the intermediate linkage have also been used in various applications such as boron neutron capture therapy for cancer treatments or antisense technologies (10,12). Boronated nucleotides were also described to display antineoplastic and hypolipidemic activities (13,14).…”

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confidence: 99%

Mechanistic Insights into the Suppression of Drug Resistance by Human Immunodeficiency Virus Type 1 Reverse Transcriptase Using α-Boranophosphate Nucleoside Analogs

Deval

Alvarez

Selmi

et al. 2005

Journal of Biological Chemistry

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A class of amino acid substitutions in drug-resistant HIV-1 reverse transcriptase (RT) is responsible for the selectively impaired incorporation of the nucleotide analog inhibitor into DNA. We have shown previously that ␣-boranophosphate nucleoside analogs suppress RT-mediated resistance when the catalytic rate is responsible for drug resistance such as in the case of K65R and dideoxy (dd)NTPs, and Q151M toward AZTTP and ddNTPs. Here, we extend this property to BH 3 -d4TTP and BH 3 -3TCTP toward their clinically relevant mutants Q151M and M184V, respectively. Pre-steady-state kinetics on mutants of the Q151M RT family reveal a 3-5-fold resistance to d4TTP. This resistance is suppressed using BH 3 -d4TTP. Likewise, resistance to 3TCTP by M184V RT (30-fold) and K65R/M184V RT (180-fold) is suppressed using BH 3 -3TCTP because of a 160-fold acceleration of the catalytic constant k pol . Mechanistic insights into the rate enhancement were obtained using various ␣-boranophosphate nucleotides. The presence of the BH 3 group renders k pol independent of amino acid substitutions present in RT. Indeed, the ϳ100-fold decrease in polymerase activity caused by the R72A substitution is restored to wild-type levels using BH 3 -dTTP. Metal ion titration studies show that ␣-boranophosphate nucleoside analogs enhance 3-8-fold the binding of Mg 2؉ ions to the active site of the RT⅐DNA⅐dNTP complex and alleviate the requirement of critical amino acids involved in phosphodiester bond formation. To our knowledge, this is the first example of rescue of polymerase activity by means of a nucleotide analog.

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Boranophosphate backbone: a mimic of phosphodiesters, phosphorothioates, and methyl phosphonates

Cited by 55 publications

References 48 publications

Thymidine Analogue Resistance Suppression by V75I of HIV-1 Reverse Transcriptase

Thymidine Analogue Resistance Suppression by V75I of HIV-1 Reverse Transcriptase

Suppression of Multidrug-resistant HIV-1 Reverse Transcriptase Primer Unblocking Activity by α-Phosphate-modified Thymidine Analogues

Mechanistic Insights into the Suppression of Drug Resistance by Human Immunodeficiency Virus Type 1 Reverse Transcriptase Using α-Boranophosphate Nucleoside Analogs

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