Enhanced subunit interactions with gemcitabine-5′-diphosphate inhibit ribonucleotide reductases
Open accessAbstract
Ribonucleotide reductases (RNRs) catalyze the conversion of nucleotides to deoxynucleotides in all organisms. The class I RNRs are composed of two subunits, α and β, with proposed quaternary structures of α2β2, α6β2, or α6β6, depending on the organism. The α subunits bind the nucleoside diphosphate substrates and the dNTP/ATP allosteric effectors that govern specificity and turnover. The β2 subunit houses the diferric Y • (1 radical per β2) cofactor that is required to initiate nucleotide reduction. 2′,2′-Difluoro-2′-deoxycytidine (F 2 C) is presently used clinically in a variety of cancer treatments and the 5′-diphosphorylated F 2 C (F 2 CDP) is a potent inhibitor of RNRs. The studies with [1′- 3 H]-F 2 CDP and [5- 3 H]-F 2 CDP have established that F 2 CDP is a substoichiometric mechanism based inhibitor (0.5 eq F 2 CDP/α) of both the Escherichia coli and the human RNRs in the presence of reductant. Inactivation is caused by covalent labeling of RNR by the sugar of F 2 CDP (0.5 eq/α) and is accompanied by release of 0.5 eq cytosine/α. Inactivation also results in loss of 40% of β2 activity. Studies using size exclusion chromatography reveal that in the E. coli RNR, an α2β2 tight complex is generated subsequent to enzyme inactivation by F 2 CDP, whereas in the human RNR, an α6β6 tight complex is generated. Isolation of these complexes establishes that the weak interactions of the subunits in the absence of nucleotides are substantially increased in the presence of F 2 CDP and ATP. This information and the proposed asymmetry between the interactions of αnβn provide an explanation for complete inactivation of RNR with substoichiometric amounts of F 2 CDP.
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KL cells respond to gemcitabine plus CHK1 inhibition.
“and to combined treatment with gemcitabine - a deoxycytidine analog that inhibits DNA synthesis and further depletes dNTP pools by targeting ribonucleotide reductase159- and CHK1 inhibitors, that abrogate the CHK1-mediated checkpoint response to replicative stress160, 161.”
From Co-occurring genomic alterations in non-small-cell lung cancer biology and therapy (2019) · cites this paper as [160]
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Cites
Nordlund, P., & Reichard, P. A. (2006) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.75.103004.142443 · Cited by 1,082 · see citing works
Action of 2',2'-difluorodeoxycytidine on DNA synthesis.
Huang, P., Chubb, S., Hertel, L. W., Grindey, G. B., & Plunkett, W. K. (1991) · PubMed · Cited by 876 · see citing works
Evaluation of the antitumor activity of gemcitabine (2',2'-difluoro-2'-deoxycytidine).
Hertel, L. W., Boder, G. B., Kroin, J. S., Rinzel, S. M., Poore, G. A., Todd, G. C., & Grindey, G. B. (1990) · PubMed · Cited by 680 · see citing works
Mackey, J. R., Mani, R. S., Selner, M., et al. (1998) · PubMed · Cited by 605 · see citing works
Structure, function, and mechanism of ribonucleotide reductases
Kolberg, M., Strand, K. R., Graff, P., & Andersson, K. K. (2004) · Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics · doi:10.1016/j.bbapap.2004.02.007 · Cited by 437 · see citing works
Inhibition of ribonucleotide reduction in CCRF-CEM cells by 2',2'-difluorodeoxycytidine.
Heinemann, V., Xu, Y.-Z., Chubb, S., et al. (1990) · Molecular Pharmacology · doi:10.1016/s0026-895x(25)09518-5 · Cited by 408 · see citing works
Determinants of resistance to 2′,2′-difluorodeoxycytidine (gemcitabine)
Bergman, A. M., Pinedo, H. M., & Peters, G. J. (2002) · Drug Resistance Updates · doi:10.1016/s1368-7646(02)00002-x · Cited by 359 · see citing works
Structure, function, and mechanism of ribonucleotide reductases
Kolberg, M. (2004) · Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics · doi:10.1016/s1570-9639(04)00054-8 · Cited by 298 · see citing works
Davidson, J. D., Ma, L., Flagella, M., Geeganage, S., Gelbert, L. M., & Slapak, C. A. (2004) · Cancer Research · doi:10.1158/0008-5472.can-03-3363 · Cited by 279 · see citing works
Baker, C. H., Banzon, J. A., Bollinger, J. M., Jr., et al. (1991) · Journal of Medicinal Chemistry · doi:10.1021/jm00110a019 · Cited by 255 · see citing works
Role of effector binding in allosteric control of ribonucleoside diphosphate reductase
Brown, N. C., & Reichard, P. A. (1969) · Journal of Molecular Biology · doi:10.1016/0022-2836(69)90056-4 · Cited by 240 · see citing works
A rapid assay for CDP reductase activity in mammalian cell extracts
Steeper, J. R., & Steuart, C. D. (1970) · Analytical Biochemistry · doi:10.1016/0003-2697(70)90092-8 · Cited by 219 · see citing works
García-Manteiga, J. M., Molina‐Arcas, M., Casado, F. J., et al. (2003) · PubMed · Cited by 212 · see citing works
Ribonucleotide reductases: radical enzymes with suicidal tendencies
Stubbe, J., & van der Donk, W. A. (1995) · Chemistry & Biology · doi:10.1016/1074-5521(95)90084-5 · Cited by 202 · see citing works
Self-citationGuittet, O., Håkansson, P., Voevodskaya, N. V., Fridd, S. L., Gräslund, A., Arakawa, H., Nakamura, Y., & Thelander, L. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m106088200 · Cited by 198 · see citing works
Bergman, A. M., Eijk, P. P., van Haperen, V. W. T. R., et al. (2005) · Cancer Research · doi:10.1158/0008-5472.can-05-0989 · Cited by 184 · see citing works
Physicochemical Characterization of Ribonucleoside Diphosphate Reductase from Escherichia coli
Thelander, L. (1973) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)43705-8 · Cited by 173 · see citing works
Thelander, L., & Larsson, B. (1976) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)33753-5 · Cited by 145 · see citing works
Amplification and purification of plasmid-encoded thioredoxin from Escherichia coli K12.
Lunn, C. A., Kathju, S., Wallace, B., Kushner, S. R., & Pigiet, V. P. (1984) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)90987-7 · Cited by 108 · see citing works
Enzymatically Active Mammalian Ribonucleotide Reductase Exists Primarily as an α6β2 Octamer
Rofougaran, R., Vodnala, M., & Hofer, A. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m605573200 · Cited by 105 · see citing works
Rompay, A. R. V., Johansson, M., & Karlsson, A. (1999) · Molecular Pharmacology · doi:10.1124/mol.56.3.562 · Cited by 103 · see citing works
Gatzeva-Topalova, P. Z., May, A. P., & Sousa, M. C. (2005) · Structure · doi:10.1016/j.str.2005.03.018 · Cited by 99 · see citing works
Kashlan, O. B., Scott, C. P., Lear, J. D., & Cooperman, B. S. (2001) · Biochemistry · doi:10.1021/bi011653a · Cited by 96 · see citing works
Seyedsayamdost, M. R., & Stubbe, J. (2006) · Journal of the American Chemical Society · doi:10.1021/ja057776q · Cited by 93 · see citing works
Self-citationShao, J., Zhou, B., Zhu, L., Qiu, W., Yuan, Y., Xi, B., & Yen, Y. (2004) · Cancer Research · doi:10.1158/0008-5472.can-03-3048 · Cited by 93 · see citing works
Chou, T. S., Heath, P. C., Patterson, L. E., et al. (1992) · Synthesis · doi:10.1055/s-1992-26167 · Cited by 87 · see citing works
Direct cloning of the trxB gene that encodes thioredoxin reductase
Russel, M., & Model, P. (1985) · Journal of Bacteriology · doi:10.1128/jb.163.1.238-242.1985 · Cited by 86 · see citing works
Excision of 2',2'-difluorodeoxycytidine (gemcitabine) monophosphate residues from DNA.
Gandhi, V. V., Legha, J., Chen, F., et al. (1996) · PubMed · Cited by 77 · see citing works
Bollinger, J. M., Jr., Hangtong, W., Ravi, N., et al. (1995) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(95)58052-2 · Cited by 73 · see citing works
van der Donk, W. A., Yu, G., Pérez, L., et al. (1998) · Biochemistry · doi:10.1021/bi9729357 · Cited by 70 · see citing works
DNA Polymerases as Targets of Anticancer Nucleosides
Miura, S., & Izuta, S. (2004) · Current Drug Targets · doi:10.2174/1389450043490578 · Cited by 66 · see citing works
Jordheim, L. P., Guittet, O., Lepoivre, M., et al. (2005) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-05-0121 · Cited by 66 · see citing works
A New Method for the Synthesis of Furanose Derivatives of Aldohexoses1
Kohn, P. M., Samaritano, R. H., & Lerner, L. M. (1965) · Journal of the American Chemical Society · doi:10.1021/ja00951a041 · Cited by 62 · see citing works
Salowe, S. P., Ator, M. A., & Stubbe, J. (1987) · Biochemistry · doi:10.1021/bi00386a024 · Cited by 59 · see citing works
Self-citationUppsten, M., Färnegårdh, M., Domkin, V., et al. (2006) · Journal of Molecular Biology · doi:10.1016/j.jmb.2006.03.035 · Cited by 59 · see citing works
Mechanistic Investigations of Ribonucleotide Reductases
Licht, S. L., & Stubbe, J. (1999) · Elsevier eBooks · doi:10.1016/b978-0-08-091283-7.00114-4 · Cited by 57 · see citing works
Self-citationSalowe, S. P., Bollinger, J. M., Jr., Ator, M. A., Stubbe, J., McCracken, J., Peisach, J., Samano, M. C., & ROBINS, M. J. (1993) · Biochemistry · doi:10.1021/bi00210a026 · Cited by 56 · see citing works
Self-citationFritscher, J., Artin, E., Wnuk, S. F., et al. (2005) · Journal of the American Chemical Society · doi:10.1021/ja043111x · Cited by 53 · see citing works
Ingemarson, R., & Thelander, L. (1996) · Biochemistry · doi:10.1021/bi960184n · Cited by 51 · see citing works
Sequence analysis of the large and small subunits of human ribonucleotide reductase
Pavloff, N., Rivard, D., Masson, S., et al. (1992) · DNA sequence · doi:10.3109/10425179209020807 · Cited by 37 · see citing works
Usova, E. V., & Eriksson, S. (1997) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1997.t01-2-00762.x · Cited by 36 · see citing works
Silva, D. J., Stubbe, J., Samano, V. A., & Robins, M. J. (1998) · Biochemistry · doi:10.1021/bi972934e · Cited by 26 · see citing works
Self-citationOrtigosa, A. D., Hristova, D., Perlstein, D. L., Zhang, Z., Huang, M., & Stubbe, J. (2006) · Biochemistry · doi:10.1021/bi0610404 · Cited by 24 · see citing works
Self-citationGemcitabine: Actions and Interactions
Plunkett, W. K., Huang, P., & Gandhi, V. V. (1997) · Nucleosides and Nucleotides · doi:10.1080/07328319708006168 · Cited by 22 · see citing works
Human M1 subunit of ribonucleotide reductase: cDNA sequence and expression in stimulated lymphocytes
Parker, N. J., Begley, C. G., & Fox, R. M. (1991) · Nucleic Acids Research · doi:10.1093/nar/19.13.3741 · Cited by 14 · see citing works
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- Lohman GJS (2006) PhD Thesis (MIT, Cambridge, MA).
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Co-occurring genomic alterations in non-small-cell lung cancer biology and therapy
Skoulidis, F., & Heymach, J. V. (2019) · Nature Reviews Cancer · doi:10.1038/s41568-019-0179-8 · Cited by 1,019 · see citing works
Avval, F. Z., & Holmgren, A. (2009) · Journal of Biological Chemistry · doi:10.1074/jbc.m809338200 · Cited by 169 · see citing works
- Authors
- Wang, J., Lohman, G. J. S., & Stubbe, J.
- Affiliations
- Massachusetts Institute of Technology (US)
- Momenta Pharmaceuticals (United States) (US)
- Corresponding author
- Stubbe, JoAnne
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2007-09-04
- Volume
- 104
- Issue
- 36
- Pages
- 14324-14329
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC1964847
- OpenAlex
- W2037851792
- Cited by
- 112
- References
- 45
- Citations by year
- 2026: 4 · 2025: 2 · 2024: 4 · 2023: 2 · 2022: 6 · 2021: 4 · 2020: 5 · 2019: 5 · 2018: 3 · 2017: 5 · 2016: 4 · 2015: 7 · 2014: 6 · 2013: 7 · 2012: 14
- FWCI
- 3.31
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Eli Lilly and Company
- Massachusetts Institute of Technology
- National Institutes of Health
- National Institute of General Medical Sciences (GM29595, R01GM029595)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
- Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- ribonucleotide reductase, subunit interactions, alpha subunit, beta2 subunit, quaternary structure, allosteric effectors, ATP, enzyme inactivation mechanism
- MeSH terms
- Gemcitabine, Cysteine, Cytidine Diphosphate, Deoxycytidine, Enzyme Activation, Escherichia coli, Humans, Molecular Weight, Protein Binding, Ribonucleotide Reductases, Time Factors, Molecular Structure, Protein Subunits
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