Genetic and enzymatic basis of hygromycin B resistance in Escherichia coli

Rao, R. Nagaraja; Allen, Norris E.; Hobbs, J. N.; Alborn, William E.; Kirst, Herbert A.; Paschal, Jonathan W.

doi:10.1128/aac.24.5.689

Cited by 88 publications

(45 citation statements)

References 27 publications

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“…Hygromycin B is an aminoglycoside antibiotic produced by Streptomyces hygroscopicus (Mann and Bromer 1958). Widely used in veterinary medicine (Kelley and Olsen 1960;Biehl 1986) and in cell culture selections (Gritz and Davies 1983;Rao et al 1983), it kills bacteria (Davies et al 1965), fungi, and higher eukaryotic cells, including mammalian cells (Gonzáles et al 1978;Eustice and Wilhelm 1984b).…”

Section: Introductionmentioning

confidence: 99%

Structural basis for hygromycin B inhibition of protein biosynthesis

Borovinskaya¹,

Shoji²,

Fredrick³

et al. 2008

RNA

105

102

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Aminoglycosides are one of the most widely used and clinically important classes of antibiotics that target the ribosome. Hygromycin B is an atypical aminoglycoside antibiotic with unique structural and functional properties. Here we describe the structure of the intact Escherichia coli 70S ribosome in complex with hygromycin B. The antibiotic binds to the mRNA decoding center in the small (30S) ribosomal subunit of the 70S ribosome and induces a localized conformational change, in contrast to its effects observed in the structure of the isolated 30S ribosomal subunit in complex with the drug. The conformational change in the ribosome caused by hygromycin B binding differs from that induced by other aminoglycosides. Also, in contrast to other aminoglycosides, hygromycin B potently inhibits spontaneous reverse translocation of tRNAs and mRNA on the ribosome in vitro. These structural and biochemical results help to explain the unique mode of translation inhibition by hygromycin B.

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Section: Introductionmentioning

confidence: 99%

Structural basis for hygromycin B inhibition of protein biosynthesis

Borovinskaya¹,

Shoji²,

Fredrick³

et al. 2008

RNA

105

102

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“…On the other hand, the corresponding residues for HygB binding were not conserved in the amino acid sequence or in the resulting structure. This might have been because the two enzymes are known to phosphorylate HygB at different sites, 11,12) and thus the binding mode of HygB to the enzymes may be different.…”

Section: Three-dimensional Modeling Of Hyg10mentioning

confidence: 99%

“…HPH and HYG show relatively low identity, of approximately 30%, and phosphorylate hygromycin B (HygB) at different sites. 11,12) The introduction of seven amino acid substitutions and a duplication of three amino acids, obtained by natural mutations, left the mutant gene (hyg10) functional as a selection marker at up to 74 C in T. thermophilus. A precise enzymatic characterization of HYG10 is also given.…”

Section: )mentioning

confidence: 99%

Directed Evolution for Thermostabilization of a Hygromycin B Phosphotransferase fromStreptomyces hygroscopicus

Sugimoto¹,

Takakura²,

Shiraki³

et al. 2013

Bioscience, Biotechnology, and Biochemistry

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“…The hpt gene, which confers resistance to the antibiotic Hygromycin B, was isolated from a plasmid found in E. coli W677 (Kuhstoss and Rao, 1983;Rao et al, 1983). Mode of action: APH4, the protein product from hpt, catalyzes the phosphorylation of the 4-hydroxyl group of Hygromycin B, rendering it biologically inactive (Pardo et al, 1985).…”

Section: Sourcementioning

confidence: 99%

Biosafety considerations for selectable and scorable markers used in cassava (Manihot esculentaCrantz) biotechnology

Petersen¹,

Umbeck²,

Hokanson

et al. 2005

Environ. Biosafety Res.

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Cassava is an important subsistence crop grown only in the tropics, and represents a major source of calories for many people in developing countries. Improvements in the areas of resistance to insects and viral diseases, enhanced nutritional qualities, reduced cyanogenic content and modified starch characteristics are urgently needed. Traditional breeding is hampered by the nature of the crop, which has a high degree of heterozygosity, irregular flowering, and poor seed set. Biotechnology has the potential to enhance crop improvement efforts, and genetic engineering techniques for cassava have thus been developed over the past decade. Selectable and scorable markers are critical to efficient transformation technology, and must be evaluated for biosafety, as well as efficiency and cost-effectiveness. In order to facilitate research planning and regulatory submission, the literature on biosafety aspects of the selectable and scorable markers currently used in cassava biotechnology is surveyed. The source, mode of action and current use of each marker gene is described. The potential for toxicity, allergenicity, pleiotropic effects, horizontal gene transfer, and the impact of these on food or feed safety and environmental safety is evaluated. Based on extensive information, the selectable marker genes nptII, hpt, bar/pat, and manA, and the scorable marker gene uidA, all have little risk in terms of biosafety. These appear to represent the safest options for use in cassava biotechnology available at this time.

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Genetic and enzymatic basis of hygromycin B resistance in Escherichia coli

Cited by 88 publications

References 27 publications

Structural basis for hygromycin B inhibition of protein biosynthesis

Structural basis for hygromycin B inhibition of protein biosynthesis

Directed Evolution for Thermostabilization of a Hygromycin B Phosphotransferase fromStreptomyces hygroscopicus

Biosafety considerations for selectable and scorable markers used in cassava (Manihot esculentaCrantz) biotechnology

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