scholarly journals Identification of Sites Influencing the Folding and Subunit Assembly of the P22 Tailspike Polypeptide Chain Using Nonsense Mutations

Genetics ◽  
1987 ◽  
Vol 117 (2) ◽  
pp. 157-171
Author(s):  
Bentley Fane ◽  
Jonathan King
Keyword(s):  
Polypeptide Chain ◽  
Acid Sites ◽  
Temperature Sensitive ◽  
Primary Defect ◽  
Subunit Assembly ◽  
Temperature Range ◽  
Lethal Phenotype ◽  
Different Temperatures ◽  
P22 Tailspike ◽  
Phage Growth

ABSTRACT Amber mutations have been isolated and mapped to more than 60 sites in gene 9 of P22 encoding the thermostable phage tailspike protein. Gene 9 is the locus of over 30 sites of temperature sensitive folding (tsf) mutations, which affect intermediates in the chain folding and subunit association pathway. The phenotypes of the amber missense proteins produced on tRNA suppressor hosts inserting serine, glutamine, tryosine and leucine have been determined at different temperatures. Thirty-three of the sites are tolerant, producing functional proteins with any of the four amino acids inserted at the sites, independent of temperature. Tolerant sites are concentrated at the N-terminal end of the protein indicating that this region is not critical for conformation or function. Sixteen of the sites yield temperature sensitive missense proteins on at least one nonsense suppressing host. Most of the sites with ts phenotypes map to the central region of the gene which is also the region where most of the tsf mutations map. Mutations at 15 of the sites have a lethal phenotype on at least one tRNA suppressor host. For nine out of ten sites tested with at least one lethal phenotype, the primary defect was in the folding or subunit association of the missense polypeptide chain. This analysis of the tailspike missense proteins distinguishes three classes of amino acid sites in the polypeptide chain; residues whose side chains contribute little to folding, subunit assembly or function; residues critical for maintaining the folding and subunit assembly pathway at the high end of the temperature range of phage growth; and residues critical over the entire temperature range of growth.

TEMPERATURE-SENSITIVE MUTANTS BLOCKED IN THE FOLDING OR SUBUNIT ASSMBLY OF THE BACTERIOPHAGE P22 TAILSPIKE PROTEIN. I. FINE-STRUCTURE MAPPING

Genetics ◽  
1980 ◽  
Vol 96 (2) ◽  
pp. 331-352 ◽  
Author(s):  
Donna H Smith ◽  
Peter B Berget ◽  
Jonathan King
Keyword(s):  
Fine Structure ◽  
Polypeptide Chain ◽  
Cell Attachment ◽  
Physical Map ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Structure Mapping ◽  
Bacteriophage P22 ◽  
P22 Tailspike ◽  
Ts Mutations

ABSTRACT As part of a study of protein folding, we have constructed a fine-structure map of 9 existing and 29 newly isolated UV- and hydroxylamine-induced temperature-sensitive (ts) mutations in gene 9 of Salmonella bacteriophage P22. Gene 9 specifies the polypeptide chain of the multimeric tail spikes, six of which form the cell attachment organelle of the phage. The 38 ts mutants were mapped against deletion lysogens with endpoints in gene 9. They mapped in 10 of the 15 deletion intervals. Two- and three-factor crosses between mutants within each interval indicated that at least 31 ts sites are represented among the 38 mutants. To determine the distribution of ts sites within the physical map, we identified the protein fragments from infection of su- hosts with 10 gene 9 amber mutants. Their molecular weights, ranging from 13,900 to 55,000 daltons, were combined with the genetic data to yield a composite map of gene 9. The 31 ts sites were distributed through most of the gene, but were most densely clustered in the central third.—None of the ts mutant pairs tested exhibited intragenic complementation. Studies of the defective phenotypes of the ts mutants (Goldenberg and King 1981; Smith and King 1981) revealed that most do not affect the thermostability of the mature protein, but instead prevent the folding or subunit assembly of the mutant chains synthesized at restrictive temperature. Thus, many of thes ts mutations identify sites in the polypeptide chain that are critical for the folding or maturation of the tail-spike protein.

scholarly journals A Genetic Analysis of Interactions with Spc110p Reveals Distinct Functions of Spc97p and Spc98p, Components of the Yeast γ-Tubulin Complex

1998 ◽  
Vol 9 (8) ◽  
pp. 2201-2216 ◽  
Author(s):  
Thu Nguyen ◽  
Dani B.N. Vinh ◽  
Douglas K. Crawford ◽  
Trisha N. Davis
Keyword(s):  
Spindle Pole Body ◽  
Spindle Pole ◽  
Amino Terminus ◽  
Temperature Sensitive ◽  
Microtubule Organizing Center ◽  
Synthetic Lethal ◽  
Lethal Phenotype ◽  
Amino Terminal ◽  
N Terminus ◽  
Two Hybrid

The spindle pole body (SPB) in Saccharomyces cerevisiae functions as the microtubule-organizing center. Spc110p is an essential structural component of the SPB and spans between the central and inner plaques of this multilamellar organelle. The amino terminus of Spc110p faces the inner plaque, the substructure from which spindle microtubules radiate. We have undertaken a synthetic lethal screen to identify mutations that enhance the phenotype of the temperature-sensitive spc110–221 allele, which encodes mutations in the amino terminus. The screen identified mutations inSPC97 and SPC98, two genes encoding components of the Tub4p complex in yeast. The spc98–63allele is synthetic lethal only with spc110 alleles that encode mutations in the N terminus of Spc110p. In contrast, thespc97 alleles are synthetic lethal withspc110 alleles that encode mutations in either the N terminus or the C terminus. Using the two-hybrid assay, we show that the interactions of Spc110p with Spc97p and Spc98p are not equivalent. The N terminus of Spc110p displays a robust interaction with Spc98p in two different two-hybrid assays, while the interaction between Spc97p and Spc110p is not detectable in one strain and gives a weak signal in the other. Extra copies of SPC98 enhance the interaction between Spc97p and Spc110p, while extra copies of SPC97interfere with the interaction between Spc98p and Spc110p. By testing the interactions between mutant proteins, we show that the lethal phenotype in spc98–63 spc110–221 cells is caused by the failure of Spc98–63p to interact with Spc110–221p. In contrast, the lethal phenotype in spc97–62 spc110–221 cells can be attributed to a decreased interaction between Spc97–62p and Spc98p. Together, these studies provide evidence that Spc110p directly links the Tub4p complex to the SPB. Moreover, an interaction between Spc98p and the amino-terminal region of Spc110p is a critical component of the linkage, whereas the interaction between Spc97p and Spc110p is dependent on Spc98p.

scholarly journals Genetic analysis of temperature-sensitive lethal mutants of Salmonella typhimurium.

Genetics ◽  
1989 ◽  
Vol 123 (4) ◽  
pp. 625-633 ◽  
Author(s):  
M B Schmid ◽  
N Kapur ◽  
D R Isaacson ◽  
P Lindroos ◽  
C Sharpe
Keyword(s):  
High Temperature ◽  
Genetic Analysis ◽  
Salmonella Typhimurium ◽  
Complex Medium ◽  
Temperature Sensitive ◽  
Nonpermissive Temperature ◽  
Anaerobic Conditions ◽  
Temperature Growth ◽  
Lethal Phenotype ◽  
Lethal Mutations

Abstract We have isolated 440 mutants of Salmonella typhimurium that show temperature-sensitive growth on complex medium at 44 degrees. Approximately 16% of the mutations in these strains have been mapped to 17 chromosomal locations; two of these chromosomal locations seem to include several essential genes. Genetic analysis of the mutations suggests that the collection saturates the genes readily mutable to a ts lethal phenotype in S. typhimurium. Physiological characteristics of the ts lethal mutants were tested: 6% of the mutants can grow at high temperature under anaerobic conditions, 17% can grow when the medium includes 0.5 M KCl, and 9% of the mutants die after a 2-hr incubation at the nonpermissive temperature. Most ts lethal mutations in this collection probably affect genes required for growth at all temperatures (not merely during high temperature growth) since Tn10 insertions that cause a temperature-sensitive lethal phenotype are rare.

The Design of AlGaN/GaN HEFT-Micro-Accelerometer and Temperature- Dependence Electrical Performance

2011 ◽  
Vol 483 ◽  
pp. 174-179 ◽  
Author(s):  
Ting Liang ◽  
Jian Jun Tang ◽  
Qian Qian Zhang ◽  
Yong Wang ◽  
Jing Li ◽  
...  
Keyword(s):  
Temperature Dependence ◽  
Electrical Performance ◽  
Saturation Current ◽  
Temperature Range ◽  
Different Temperatures ◽  
Piezoresistance Coefficient ◽  
Micro Accelerometer

In this paper, We use a novel principle to detect acceleration and report how I-V characteristics and piezoresistance coefficient of AlGaN/GaN HEFT-micro-accelerometer are affected by setting different temperatures. It is shown that saturation current of device would go down if the temperature goes up, which is about 0.028mA/°C, based on the research. However, the device can work well at the temperature range of -50°C to 50°C, which indicates that it can work safely in the larger temperature range.

scholarly journals A Study on the Effect of Ultrasonic Treatment on the Microstructure of Sn-30 wt.% Bi Alloy

Materials ◽  
2018 ◽  
Vol 11 (10) ◽  
pp. 1870
Author(s):  
Shuo Wang ◽  
Jinwu Kang ◽  
Xiaopeng Zhang ◽  
Zhipeng Guo
Keyword(s):  
Ultrasonic Treatment ◽  
Acoustic Streaming ◽  
Eutectic Structure ◽  
Eutectic Phase ◽  
Ultrasonic Cavitation ◽  
Mixed Effect ◽  
Temperature Range ◽  
Different Temperatures

The effect of ultrasonic treatment on the microstructure of Sn-30 wt.% Bi alloy was studied at different temperatures. Results showed that the ultrasonic treatment could effectively refine the microstructure of Sn-30 wt.% Bi alloy at a temperature range between the liquidus and solidus. Application of the ultrasound could fragment the primary Sn dendrites during solidification due to a mixed effect of ultrasonic cavitation and acoustic streaming. The divorced eutectic formed when the ultrasonic treatment was applied for the whole duration of the solidification. The eutectic phase grew and surrounded the primary Sn dendrite, and pure Bi phase grew in between the Sn dendritic fragments. The mechanism of the fragmentation of dendrites and the divorced eutectic structure by ultrasonic treatment was discussed.

scholarly journals Application of headspace for research volatile organic compounds emitted from building materials

2018 ◽  
Vol 28 ◽  
pp. 01019
Author(s):  
Beata Kultys ◽  
Karolina Waląg
Keyword(s):  
Volatile Organic Compounds ◽  
Organic Compounds ◽  
Building Materials ◽  
Heating Temperature ◽  
Mineral Wool ◽  
Chromatography Method ◽  
Temperature Range ◽  
Volatile Organic ◽  
Different Temperatures

Headspace technique and gas chromatography method with mas detector has been used for the determination of volatile organic compounds (VOC) emitted from various building and finishing materials, such as sealing foams, mounting strips, paints, varnishes, floor coverings. The tests were carried out for different temperatures (in the temperature range of 60 to 180 °C) and the time of heated vials with tested materials inside. These tests were conducted to verify the possibility of use this method of determination the VOC emission. Interpretation of chromatograms and mass spectra allowed to identify the type of compounds emitted from the tested materials and the optimum time and temperature for each type of material was determined. The increase in heating temperature of the samples resulted in increase the type and number of identified compounds: for four materials the increase was in the whole temperature range, for others it was from 90 °C. On the other hand, emission from mineral wool was low in whole temperature range. 30-minutes heating of the samples was sufficient to identify emitted compounds for most of tested materials. Applying a longer time, i.e. 24 hours, significantly increased the sensitivity of the method.

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