Frameshift mutation in the lysozyme gene of bacteriophage T4: Demonstration of the insertion of five bases, and a summary of in vivo codons and lysozyme activities

Ocada, Yoshiko; Amagase, Shizuko; Tsugita, Akira

doi:10.1016/0022-2836(70)90428-6

Cited by 32 publications

(3 citation statements)

References 30 publications

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“…1 and 2. The guanido group of Arg 145 also extends across the mouth of the opening to within about 4 A of the carboxyl of Glu 21, although the two groups do not obviously form a salt link.…”

Section: Description Of the Structurementioning

confidence: 99%

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The Three Dimensional Structure of the Lysozyme from Bacteriophage T4

Matthews

Remington

1974

Proc. Natl. Acad. Sci. U.S.A.

170

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The three dimensional structure of the lysozyme from bacteriophage T4 has been determined from a 2.5 A resolution electron density map. About 60% of the molecule is in a helical conformation and there is one region consisting of antiparallel (-structure. The polypeptide backbone folds into two' distinct lobes linked in part by a long helix. In the region between the two lobes, there is a cleft which deepens into a hole or cavity, about 6-8 A in diameter, extending'from one side of the molecule to the other. This opening is closed off by side chains which extend to within 3-5 A of each other. A number of mutant lysozymes in which residues in the vicinity of the opening are modified have markedly reduced catalytic activity, suggesting that this region of the molecule may be catalytically important. The three dimensional structure of -T4 phage lysozyme is quite different from that of hen egg-white lysozyme although it is not clear at this time whether or not the mechanisms of catalysis of the respective enzymes are related.T4 phage lysozyme is an enzyme produced in cells of Escherichia coli after infection with bacteriophage T4. The enzyme has similar catalytic activity to that of hen egg-white lysozyme, both being endoacetylmuramidases (1). The molecular weight of the enzyme is 18,700, and the amino-acid sequence has been determined (2). Furthermore, a number of lysozymes have been isolated from mutant strains carrying frame-shift or amber mutations in the phage genome and have been used to demonstrate in vivo certain features of the genetic code (3-5).In this preliminary report, we describe the three dimensional structure of T4 phage lysozyme as determined by x-ray crystallography from a 2.5 A resolution electron density map. Additional details will be given in a subsequent publication. CrystallizationThe protein was purified using essentially the method of Tsugita et al.(1) except that 1 mM mercaptoethanol was added to all buffers. Conditions for obtaining the crystals have been described previously (6). Before x-ray photography the crystals were equilibrated with a standard mother liquor consisting of 1.05 M K2HPO4, 1.26 M NaH2PO4, 0.23 M NaCl, 1.4 mM mercaptoethanol, pH 6.7. In this solution the crystals float.The crystals have space group P3221 with cell dimensions a = b = 61.1 A, c = 96.3 A and one molecule in each of the six asymmetric units. Data collectionDiffraction data were recorded photographically using conventional Buerger precession cameras and integrated intensities measured with a computer controlled drum film scanner (7). Sets of 12 films sufficed to measure 92% of the data to 2.5 A and 53% of the data between 2.4 A and 2.5 A. With the films used, the Bijvoet differences were obtained for about half the reflections. Systematic errors in the measurement of the Friedel pairs were reduced by a method of local scaling to be described elsewhere (ref 8, B. W. Matthews et al., manuscript in preparation). Altogether, about 27,000 intensities were measured for the parent crystals and the two heav...

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Section: Description Of the Structurementioning

confidence: 99%

“…The molecular weight of the enzyme is 18,700, and the amino-acid sequence has been determined (2). Furthermore, a number of lysozymes have been isolated from mutant strains carrying frame-shift or amber mutations in the phage genome and have been used to demonstrate in vivo certain features of the genetic code (3)(4)(5).…”

mentioning

confidence: 99%

The Three Dimensional Structure of the Lysozyme from Bacteriophage T4

Matthews

Remington

1974

Proc. Natl. Acad. Sci. U.S.A.

170

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“…If A--G and C--U replacements were to occur exclusively at the third position of the codon, the DNA of E. coli could vary from 31% G +C content to 68% G +C content without any change in the amino acids of any protein (Singer and Ames, 1970). By identifying some of the codons utilized in vivo in the wild type T4 lysozyme, Streisinger et al (1966) and Okada et al (1970) have proposed that the base composition of the DNA reflects the base composition at the third position of the codon. Relatively low intramolecular heterogeneity in nucleotide composition in AT or GC rich bacteria may be expected if one supposes that AT rich bacterial species are likely to use genetic codons containing A or U at the third coding positions and GC rich bacterial species use genetic codons containing G or C at the third coding positions.…”

Section: Dna Fragments Aboutmentioning

confidence: 99%

Nucleotide distribution in bacterial DNA's differing in G + C content

Yamagishi

1974

J Mol Evol

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Summary. The DNA's of Micrococcus lysodeikticus and Clostridium per/ringenswere fragmented to about 7 000 nucleotide pairs long by shear and fractionated with respect to buoyant density of mercury complexes in Cs2SO 4. The distribution of G + C content in both DNA's was characteristically asymmetric. In M. lysodeikticus DNA, low G + C fragments were more numerous than high G + C fragments, whereas in C. per/ringens DNA, high G + C fragments were more numerous than low G + C fragments. The G+ C content of fragments of M. lysodeikticus DNA varied from 70 to 77 %, with a mean and standard deviation of 73.7 ± 1.92 % G + C and that of C. per/ringens DNA varied from 27 to 34 %, with a mean and standard deviation of 29.8 i t .34 % G + C. Tile standard deviation was smaller than that of Escherichia coli DNA fragments of similar size. Biological meanings of relatively low heterogeneity in nucleotide composition in M. lysodeikticus and C. per/ringens are discussed. Key words: Micrococcus lysodeikticus --Clostridium per/ringens --DNA --Hg --Cs2SO 4 --Low Heterogeneity --Composition Bias at the Third Coding Positions --tRNA.It was recently reported (Yamagishi, t970; Yamagishi and Takahashi, 197t) that the bacterial DNA's of Escherichia coli and Bacillus subtilis contained segments of dissimilar nucleotide composition. In this communication, the result of a similar investigation carried out with bacterial DNA's having extreme G-k C composition is presented.Clostridium per/ringens strain PB6K and Micrococcus lysodeikticus strain 2665 used in the present investigation were obtained from Research Institute of Microbial Diseases, Osaka University, Japan. M. lysodeikticus was grown for 24 hrs in low P-k-broth (pH 7.2) containing per liter 10 g low P-polypeptone (Inorganic phosphorus content decreased to 0.008 % in our laboratory) and 2.5 g NaCI. C. per/ringens was

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Structure and Function of RNA Bacteriophages

Fiers¹

1979

Comprehensive Virology Volume 13: Structure and Assembly

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Frameshift mutation in the lysozyme gene of bacteriophage T4: Demonstration of the insertion of five bases, and a summary of in vivo codons and lysozyme activities

Cited by 32 publications

References 30 publications

The Three Dimensional Structure of the Lysozyme from Bacteriophage T4

The Three Dimensional Structure of the Lysozyme from Bacteriophage T4

Nucleotide distribution in bacterial DNA's differing in G + C content

Structure and Function of RNA Bacteriophages

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