kanamycin kinase


Summary: A class of enzymes that inactivate aminocyclitol-aminoglycoside antibiotics (AMINOGLYCOSIDES) by regiospecific PHOSPHORYLATION of the 3' and/or 5' hydroxyl.

Top Publications

  1. Boehr D, Lane W, Wright G. Active site labeling of the gentamicin resistance enzyme AAC(6')-APH(2") by the lipid kinase inhibitor wortmannin. Chem Biol. 2001;8:791-800 pubmed
  2. Burk D, Hon W, Leung A, Berghuis A. Structural analyses of nucleotide binding to an aminoglycoside phosphotransferase. Biochemistry. 2001;40:8756-64 pubmed
    ..This alternate binding mode is likely a conserved feature among aminoglycoside kinases and could be exploited for the structure-based drug design of compounds to combat antibiotic resistance...
  3. Boehr D, Thompson P, Wright G. Molecular mechanism of aminoglycoside antibiotic kinase APH(3')-IIIa: roles of conserved active site residues. J Biol Chem. 2001;276:23929-36 pubmed
    ..These results lay the foundation for the synthesis of transition state mimics that could reverse aminoglycoside antibiotic resistance in vivo. ..
  4. Shakya T, Wright G. Nucleotide selectivity of antibiotic kinases. Antimicrob Agents Chemother. 2010;54:1909-13 pubmed publisher
    ..These differences reveal likely different pathways in antibiotic resistance enzyme evolution and can be exploited in selective inhibitor design to counteract resistance. ..
  5. Wright G, Thompson P. Aminoglycoside phosphotransferases: proteins, structure, and mechanism. Front Biosci. 1999;4:D9-21 pubmed
    ..In particular, the revelation of structural and mechanistic similarities between APHs and Ser/Thr and Tyr kinases has set the stage for future inhibition studies which could prove important in reversing aminoglycoside resistance. ..
  6. Hallmann A, Wodniok S. Swapped green algal promoters: aphVIII-based gene constructs with Chlamydomonas flanking sequences work as dominant selectable markers in Volvox and vice versa. Plant Cell Rep. 2006;25:582-91 pubmed
    ..This strategy might be useful for all investigators that intend to transform species in which genomic sequences are not available, but sequences from related organisms exist...
  7. Okazaki A, Avison M. Aph(3')-IIc, an aminoglycoside resistance determinant from Stenotrophomonas maltophilia. Antimicrob Agents Chemother. 2007;51:359-60 pubmed
    ..Disruption of aph(3')-IIc in K279a results in decreased MICs of these drugs. ..
  8. Thompson P, Hughes D, Wright G. Mechanism of aminoglycoside 3'-phosphotransferase type IIIa: His188 is not a phosphate-accepting residue. Chem Biol. 1996;3:747-55 pubmed
    ..Positional isotope exchange experiments are consistent with direct attack of the aminoglycoside hydroxyl group on the gamma-phosphate of ATP. ..
  9. Jakobiak T, Mages W, Scharf B, Babinger P, Stark K, Schmitt R. The bacterial paromomycin resistance gene, aphH, as a dominant selectable marker in Volvox carteri. Protist. 2004;155:381-93 pubmed
    ..This makes the engineered bacterial aphH gene an efficient dominant selection marker for the transformation and co-transformation of a broad range of V. carteri strains without the recurring need for using auxotrophic recipient strains...

More Information


  1. Kim C, Haddad J, Vakulenko S, Meroueh S, Wu Y, Yan H, et al. Fluorinated aminoglycosides and their mechanistic implication for aminoglycoside 3'-phosphotransferases from Gram-negative bacteria. Biochemistry. 2004;43:2373-83 pubmed
    ..These findings collectively argue that the Gram-negative APH(3')s show significant nucleophilic participation in the transition state for the phosphate transfer reaction. ..
  2. Toth M, Chow J, Mobashery S, Vakulenko S. Source of phosphate in the enzymic reaction as a point of distinction among aminoglycoside 2''-phosphotransferases. J Biol Chem. 2009;284:6690-6 pubmed publisher
    ..These nucleotide triphosphate preferences should have ramifications for understanding of the evolution, selection, and dissemination of the genes for these important resistance enzymes. ..
  3. Kunz C, Narangajavana J, Jakowitsch J, Park Y, Delon T, Kovarik A, et al. Studies on the effects of a flanking repetitive sequence on the expression of single-copy transgenes in Nicotiana sylvestris and in N. sylvestris-N. tomentosiformis hybrids. Plant Mol Biol. 2003;52:203-15 pubmed
    ..3 dispersed repeat family on transgene expression was negligible. The silencing phenomena observed with the three single-copy transgene loci are discussed in the context of other possible triggers of silencing. ..
  4. Fong D, Berghuis A. Substrate promiscuity of an aminoglycoside antibiotic resistance enzyme via target mimicry. EMBO J. 2002;21:2323-31 pubmed publisher
    ..These results suggest a potential strategy for drug design aimed at circumventing antibiotic resistance...
  5. Thompson C, Gray G. Nucleotide sequence of a streptomycete aminoglycoside phosphotransferase gene and its relationship to phosphotransferases encoded by resistance plasmids. Proc Natl Acad Sci U S A. 1983;80:5190-4 pubmed
    ..fradiae aph with the aph genes encoded by bacterial transposons Tn5 and Tn903 showed significant nucleotide and amino acid homologies which indicated a common evolutionary origin for these antibiotic-resistance genes. ..
  6. Fojtova M, Van Houdt H, Depicker A, Kovarik A. Epigenetic switch from posttranscriptional to transcriptional silencing is correlated with promoter hypermethylation. Plant Physiol. 2003;133:1240-50 pubmed
    ..The results suggest that a switch between posttranscriptional and transcriptional gene silencing could be a mechanism leading to irrevocable shut down of gene expression within a finite number of generations. ..
  7. Kim C, Cha J, Yan H, Vakulenko S, Mobashery S. Hydrolysis of ATP by aminoglycoside 3'-phosphotransferases: an unexpected cost to bacteria for harboring an antibiotic resistance enzyme. J Biol Chem. 2006;281:6964-9 pubmed
    ..The implication is that, in the absence of selection, bacteria harboring an enzyme that catalyzes the consumption of key metabolites could experience the loss of the plasmid that encodes for the given enzyme. ..
  8. Nurizzo D, Shewry S, Perlin M, Brown S, Dholakia J, Fuchs R, et al. The crystal structure of aminoglycoside-3'-phosphotransferase-IIa, an enzyme responsible for antibiotic resistance. J Mol Biol. 2003;327:491-506 pubmed
    ..These results give new information on the basis of aminoglycoside recognition, and on the relationship between this phosphotransferase family and the protein kinases. ..
  9. Boehr D, Draker K, Koteva K, Bains M, Hancock R, Wright G. Broad-spectrum peptide inhibitors of aminoglycoside antibiotic resistance enzymes. Chem Biol. 2003;10:189-96 pubmed
    ..These peptides represent the first example of broad-spectrum inhibitors of aminoglycoside resistance enzymes. ..
  10. Smith C, Baker E. Aminoglycoside antibiotic resistance by enzymatic deactivation. Curr Drug Targets Infect Disord. 2002;2:143-60 pubmed
    ..This review focuses on the structural details of the enzymes whose crystal structures are known and on the implications of these findings for devising novel strategies to overcome resistance to this broad class of antibiotics. ..
  11. Sizova I, Fuhrmann M, Hegemann P. A Streptomyces rimosus aphVIII gene coding for a new type phosphotransferase provides stable antibiotic resistance to Chlamydomonas reinhardtii. Gene. 2001;277:221-9 pubmed
    ..We conclude that the aphVIII gene is a useful marker for nuclear transformation and promoter tagging of C. reinhardtii wild-type and probably other green algae. ..
  12. Franceschini I, Feigenbaum Lacombe V, Casanova P, Lopez Lastra M, Darlix J, Dalcq M. Efficient gene transfer in mouse neural precursors with a bicistronic retroviral vector. J Neurosci Res. 2001;65:208-19 pubmed
  13. Hon W, McKay G, Thompson P, Sweet R, Yang D, Wright G, et al. Structure of an enzyme required for aminoglycoside antibiotic resistance reveals homology to eukaryotic protein kinases. Cell. 1997;89:887-95 pubmed
    ..These results indicate that APH(3') enzymes and eukaryotic-type protein kinases share a common ancestor. ..
  14. Lerche K, Hallmann A. Stable nuclear transformation of Gonium pectorale. BMC Biotechnol. 2009;9:64 pubmed publisher
  15. Padilla I, Burgos L. Aminoglycoside antibiotics: structure, functions and effects on in vitro plant culture and genetic transformation protocols. Plant Cell Rep. 2010;29:1203-13 pubmed publisher
  16. Wieninger S, Serpersu E, Ullmann G. ATP binding enables broad antibiotic selectivity of aminoglycoside phosphotransferase(3')-IIIa: an elastic network analysis. J Mol Biol. 2011;409:450-65 pubmed publisher
    ..Additionally, we must consider correlated motions of dynamic protein domains, which show that not only connectivity but also the overall protein architecture is important for protein dynamics. ..
  17. Lallemand P, Leban N, Kunzelmann S, Chaloin L, Serpersu E, Webb M, et al. Transient kinetics of aminoglycoside phosphotransferase(3')-IIIa reveals a potential drug target in the antibiotic resistance mechanism. FEBS Lett. 2012;586:4223-7 pubmed publisher
    ..A good strategy to prevent the inactivation of aminoglycosides would be to develop uncompetitive inhibitors that interact with this key ADP-enzyme complex. ..
  18. Lee K, Hopkins J, Syvanen M. Evolved neomycin phosphotransferase from an isolate of Klebsiella pneumoniae. Mol Microbiol. 1991;5:2039-46 pubmed
    ..Our hypothesis to explain these results is that aphA1-IAB evolved under the selective pressure of neomycin use in relatively recent times...
  19. Fong D, Berghuis A. Structural basis of APH(3')-IIIa-mediated resistance to N1-substituted aminoglycoside antibiotics. Antimicrob Agents Chemother. 2009;53:3049-55 pubmed publisher
  20. Thompson P, Boehr D, Berghuis A, Wright G. Mechanism of aminoglycoside antibiotic kinase APH(3')-IIIa: role of the nucleotide positioning loop. Biochemistry. 2002;41:7001-7 pubmed
    ..The APH NPL therefore acts as a lever, promoting phosphoryl transfer to the aminoglycoside substrate, with the biological outcome of clinically relevant antibiotic resistance. ..
  21. Lopez Lastra M, Gabus C, Darlix J. Characterization of an internal ribosomal entry segment within the 5' leader of avian reticuloendotheliosis virus type A RNA and development of novel MLV-REV-based retroviral vectors. Hum Gene Ther. 1997;8:1855-65 pubmed
  22. Kovarik A, Van Houdt H, Holy A, Depicker A. Drug-induced hypomethylation of a posttranscriptionally silenced transgene locus of tobacco leads to partial release of silencing. FEBS Lett. 2000;467:47-51 pubmed
    ..Models for the possible role of DNA methylation in PTGS are discussed. ..
  23. Ferretti J, Gilmore K, Courvalin P. Nucleotide sequence analysis of the gene specifying the bifunctional 6'-aminoglycoside acetyltransferase 2"-aminoglycoside phosphotransferase enzyme in Streptococcus faecalis and identification and cloning of gene regions specifying the two activitie. J Bacteriol. 1986;167:631-8 pubmed
    ..These data suggest that the gene specifying the AAC(6')-APH(2") resistance enzyme arose as a result of a gene fusion...
  24. Berthold P, Schmitt R, Mages W. An engineered Streptomyces hygroscopicus aph 7" gene mediates dominant resistance against hygromycin B in Chlamydomonas reinhardtii. Protist. 2002;153:401-12 pubmed
    ..This advocates the hygromycin-resistance plasmid, pHyg3, as a new versatile tool for the transformation of a broad range of C. reinhardtii strains without the sustained need for using auxotrophic mutants as recipients. ..
  25. Fong D, Xiong B, Hwang J, Berghuis A. Crystal structures of two aminoglycoside kinases bound with a eukaryotic protein kinase inhibitor. PLoS ONE. 2011;6:e19589 pubmed publisher
    ..Our results provide evidence that an inhibitor for a subset of APHs can be developed in order to curtail resistance to aminoglycosides. ..
  26. Chamberlain J, Schwarze U, Wang P, Hirata R, Hankenson K, Pace J, et al. Gene targeting in stem cells from individuals with osteogenesis imperfecta. Science. 2004;303:1198-201 pubmed
  27. Chen L, Xie Z, Teng Y, Wang M, Shi M, Qian L, et al. Highly efficient selection of the stable clones expressing antibody-IL-2 fusion protein by a dicistronic expression vector containing a mutant neo gene. J Immunol Methods. 2004;295:49-56 pubmed
    ..Combined with an amplification system, this vector may have potential usage for the expression of recombinant antibody and the yield may be further improved by this expression system. ..
  28. Sauer B. Manipulation of transgenes by site-specific recombination: use of Cre recombinase. Methods Enzymol. 1993;225:890-900 pubmed
  29. Wu C, Janssen G. Translation of vph mRNA in Streptomyces lividans and Escherichia coli after removal of the 5' untranslated leader. Mol Microbiol. 1996;22:339-55 pubmed
    ..coli and S. lividans. ..
  30. Liu Y, Wu Q, Cui H, Li Q, Zhao Y, Luo J, et al. Expression of EGFP and NPTII protein is not associated with organ abnormalities in deceased transgenic cloned cattle. Cloning Stem Cells. 2008;10:421-8 pubmed publisher
    ..All data indicated that the expression of EGFP and NPTII is not associated with organ abnormalities in transgenic cloned cattle. ..
  31. Fiering S, Epner E, Robinson K, Zhuang Y, Telling A, Hu M, et al. Targeted deletion of 5'HS2 of the murine beta-globin LCR reveals that it is not essential for proper regulation of the beta-globin locus. Genes Dev. 1995;9:2203-13 pubmed
    ..In addition, this result emphasizes that when using homologous recombination to analyze complex regulatory elements in vivo, the inserted selectable marker must be removed to avoid influencing the phenotype of the mutation. ..
  32. Jia Z, Sun Y, Yuan L, Tian Q, Luo K. The chitinase gene (Bbchit1) from Beauveria bassiana enhances resistance to Cytospora chrysosperma in Populus tomentosa Carr. Biotechnol Lett. 2010;32:1325-32 pubmed publisher
    ..Fr. Similarly, Bbchit1 overexpression enhanced disease resistance to C. chrysosperma in the transformed poplar plants, indicating that is gene is potentially useful to protect the trees against fungal diseases. ..
  33. Drews R, Kolker M, Moran C, Sachar D, Chan V, Schnipper L. Genetic analysis of adenovirus E1A: induction of genetic instability and altered cell morphologic and growth characteristics are segregatable functions. Mutat Res. 1998;421:9-25 pubmed
  34. Taniguchi T, Kurita M, Ohmiya Y, Kondo T. Agrobacterium tumefaciens-mediated transformation of embryogenic tissue and transgenic plant regeneration in Chamaecyparis obtusa Sieb. et Zucc. Plant Cell Rep. 2005;23:796-802 pubmed
    ..The integration of the genes into the genome of regenerated plantlets was confirmed by Southern blot analysis. ..
  35. Briskin M, Hsu R, Boggs T, Schultz J, Rishell W, Bosselman R. Heritable retroviral transgenes are highly expressed in chickens. Proc Natl Acad Sci U S A. 1991;88:1736-40 pubmed
    ..The REV vector system as applied to the chicken represents a departure from the long-established paradigm of retroviral transgenes in mice and provides a new approach to the study of avian biology. ..
  36. Yao Y, Xiong J, Wang X, Li J, Hu Y. [The presentation of neo gene in chimeric mice obtained from ES cells]. Yi Chuan Xue Bao. 2001;28:1116-9 pubmed
    ..PCR was carried out to analyze the presentation of neo gene in tissues of chimeric mice. The results showed that many tissues harbored the neo gene, including skin, liver and blood et al. ..
  37. Yuan C, Kent C. Identification of critical residues of choline kinase A2 from Caenorhabditis elegans. J Biol Chem. 2004;279:17801-9 pubmed
  38. Gao C, Zhu X, Zhou X, Yu F, Lu B, Huang P. [Construction of eukaryotic expression vector using neomycin-resistance gene mutant as selectable marker]. Sheng Wu Gong Cheng Xue Bao. 2002;18:308-12 pubmed
    ..As a result, the expression positive ratio reaches to approximate 95% and the ratio of high expression colonies is apparently higher than the controls. ..
  39. Randhawa G, Singh M. Multiplex, construct-specific, and real-time PCR-based analytical methods for Bt rice with cry1Ac gene. J AOAC Int. 2012;95:186-94 pubmed
    ..They could also address consumers' concerns and legal disputes that may arise. ..
  40. Marinho C, Silva N, Pombo S, Santos T, Monteiro R, Gonçalves A, et al. Echinoderms from Azores islands: an unexpected source of antibiotic resistant Enterococcus spp. and Escherichia coli isolates. Mar Pollut Bull. 2013;69:122-7 pubmed publisher
    ..The data recovered are essential to improve knowledge about the dissemination of resistant strains through marine ecosystems and the possible implications involved in transferring these resistances either to other animals or to humans. ..
  41. Smith A, Awuah E, Capretta A, Brennan J. A matrix-assisted laser desorption/ionization tandem mass spectrometry method for direct screening of small molecule mixtures against an aminoglycoside kinase. Anal Chim Acta. 2013;786:103-10 pubmed publisher
    ..Inhibition constants were obtained for the aforementioned inhibitors using the MALDI-MS/MS assay, revealing several low to mid micromolar "hits", and highlighting the quantitative nature of the assay. ..
  42. Zhou H, Zhang F, Ma Z, Huang H, Jiang L, Cai T, et al. Folate polyglutamylation is involved in chromatin silencing by maintaining global DNA methylation and histone H3K9 dimethylation in Arabidopsis. Plant Cell. 2013;25:2545-59 pubmed publisher
    ..Our study makes an important contribution to understanding the complex interplay among metabolism, development, and epigenetic regulation. ..
  43. Dhekney S, Li Z, Dutt M, Gray D. Agrobacterium-mediated transformation of embryogenic cultures and plant regeneration in Vitis rotundifolia Michx. (muscadine grape). Plant Cell Rep. 2008;27:865-72 pubmed publisher
    ..Transgenic plants exhibited uniform GFP expression in cells of all plant tissues and organs including leaves, stems, roots, inflorescences and the embryo and endosperm of developing berries. ..
  44. Sangaev S, Trifonova E, Titov S, Romanova A, Kolodiazhnaia I, Komarova M, et al. [Effective expression of the gene encoding an extracellular ribonuclease of Zinnia elegans in the SR1 Nicotiana tabacum plants]. Genetika. 2007;43:1002-5 pubmed
    ..Primary tobacco transformants were characterized by a high level of RNase activity. ..
  45. Fuchs M, Chirco E, Gonsalves D. Movement of coat protein genes from a commercial virus-resistant transgenic squash into a wild relative. Environ Biosafety Res. 2004;3:5-16 pubmed
    ..This is the first report on transgene dissemination from a transgenic crop that exhibits disease resistance and hybridizes with a wild plant species without loss of fertility. ..
  46. Manickavasagam M, Ganapathi A, Anbazhagan V, Sudhakar B, Selvaraj N, Vasudevan A, et al. Agrobacterium-mediated genetic transformation and development of herbicide-resistant sugarcane (Saccharum species hybrids) using axillary buds. Plant Cell Rep. 2004;23:134-43 pubmed
    ..These results show that, with this protocol, generation and multiplication of transgenic shoots can be achieved in about 5 months with transformation efficiencies as high as 50%...
  47. Sizova I, Hegemann P, Furmann M, Danilenko V. [Streptomyces rimosus aminoglycoside-3'-phosphotransferase VIII: comparisons with aminoglycoside-3'-phosphotransferases of aminoglycoside-producing strains with eukaryotic protein kinases]. Mol Biol (Mosk). 2002;36:27-36 pubmed
    ..APHVIII differed from aminoglycoside 3'-phosphotransferases of aminoglycoside-producing actinomycete strains and of clinical isolates, and can be classed to a separate group. ..
  48. Dunbar C, Seidel N, Doren S, Sellers S, Cline A, Metzger M, et al. Improved retroviral gene transfer into murine and Rhesus peripheral blood or bone marrow repopulating cells primed in vivo with stem cell factor and granulocyte colony-stimulating factor. Proc Natl Acad Sci U S A. 1996;93:11871-6 pubmed
  49. Dan Y, Yan H, Munyikwa T, Dong J, Zhang Y, Armstrong C. MicroTom--a high-throughput model transformation system for functional genomics. Plant Cell Rep. 2006;25:432-41 pubmed
    ..The utility of this MicroTom model transformation system for functional genomic studies, such as identification of genes related to important agricultural traits and gene function, is discussed. ..
  50. Tang W. Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.). Cell Res. 2001;11:237-43 pubmed
    ..The successful expression of the GUS gene in different families of loblolly pine suggests that this transformation system is probably useful for the production of the genetically modified conifers. ..
  51. Steinecke P, Herget T, Schreier P. Expression of a chimeric ribozyme gene results in endonucleolytic cleavage of target mRNA and a concomitant reduction of gene expression in vivo. EMBO J. 1992;11:1525-30 pubmed
    ..However, the mutant ribozyme--target complex was more stable than the wildtype ribozyme--target complex. This may result in an overestimation of the antisense effect contributing to the overall reduction of gene expression. ..
  52. Cunningham L, Su C. Astrocyte delivery of glial cell line-derived neurotrophic factor in a mouse model of Parkinson's disease. Exp Neurol. 2002;174:230-42 pubmed
    ..These results indicate that continuous exposure to low levels of GDNF provided by transgenic astrocytes provides marked neuroprotection of nigral dopaminergic neurons. (c)2002 Elsevier Science (USA). ..
  53. Wang P, Wang Q, Sims P, Hyde J. Rapid positive selection of stable integrants following transfection of Plasmodium falciparum. Mol Biochem Parasitol. 2002;123:1-10 pubmed