A Point Mutation Increasing the Stability of Human Phosphoglucose Isomerase

Gelsolin modulates the actin cytoskeleton in the cytoplasm and clears the circulation of stray filaments. In vitro, gelsolin cleaves, nucleates and caps actin filaments, activities that are calcium-dependent. Both cellular and secreted forms share a sequence of 730 residues comprising six homologous modules termed G1–G6. A disulphide bond is formed in secreted G2, whereas in the cytoplasm it remains reduced. A point mutation in G2 causes an amyloidosis with neurological, ophthalmological and dermatological symptoms. This mutation does not affect the cytoplasmic form, while the secreted form is proteolysed. As a first step towards understanding how gelsolin folds and functions in different cellular compartments, we have characterized at equilibrium the urea-induced unfolding of G1 and G2, with or without calcium and/or disulphide bond. G1 and G2 both exhibit two-state unfolding behaviour and are stabilized by calcium. The disulphide bond also contributes to the stability of G2. In the absence of Ca2+ and disulphide bond, G2 adopts a non-native conformation, suggesting that folding of G2 in the cytoplasm relies on the presence of surrounding modules or other molecular partners.

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Recurrence of repeat-induced point mutation (RIP) in Neurospora crassa.

Genetics ◽

10.1093/genetics/127.4.699 ◽

1991 ◽

Vol 127 (4) ◽

pp. 699-710 ◽

Cited By ~ 3

Author(s):

E B Cambareri ◽

M J Singer ◽

E U Selker

Keyword(s):

Neurospora Crassa ◽

Point Mutation ◽

Dna Sequences ◽

High Frequency ◽

First Generation ◽

Vegetative Cells ◽

Homologous Sequences ◽

Sequences Analysis ◽

The Stability ◽

Repeat Induced Point Mutation

Abstract Duplicate DNA sequences in the genome of Neurospora crassa can be detected and mutated in the sexual phase of the life cycle by a process termed RIP (repeat-induced point mutation). RIP occurs in the haploid nuclei of fertilized, premeiotic cells before fusion of the parental nuclei. Both copies of duplications of gene-sized sequences are affected in the first generation at frequencies of approximately 50-100%. We investigated the extent to which sequences altered by RIP remain susceptible to this process in subsequent generations. Duplications continued to be sensitive to RIP, even after six generations. The fraction of progeny showing evidence of RIP decreased rapidly, however, apparently as a function of the extent of divergence of the duplicated sequences. Analysis of the stability of heteroduplexes of DNA altered by RIP and their native counterpart indicated that linked duplications diverged further than did unlinked duplications. DNA methylation, a common feature of sequences altered by RIP, did not seem to inhibit the process. A sequence that had become resistant to RIP was cloned and reintroduced into Neurospora in one or more copies to investigate the basis of the resistance. The altered sequence regained its methylation in vegetative cells, indicating that the methylation of sequences altered by RIP observed in vegetative cells is a consequence of the mutations. Duplication of the sequence restored its sensitivity to RIP suggesting that resistance to the process was due to loss of similarity between the duplicated sequences. Consistent with this, we found that the resistant sequence did not trigger RIP of the native homologous sequences of the host, even when no other partner was available. High frequency intrachromatid recombination, which is temporally associated with RIP, was more sensitive than RIP to alterations in the interacting sequences.

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A single point mutation (Glu85Arg) increases the stability of the thioredoxin from Escherichia coli

Protein Engineering Design and Selection ◽

10.1093/protein/14.4.255 ◽

2001 ◽

Vol 14 (4) ◽

pp. 255-260 ◽

Cited By ~ 19

Author(s):

E. Pedone ◽

M. Saviano ◽

M. Rossi ◽

S. Bartolucci

Keyword(s):

Escherichia Coli ◽

Point Mutation ◽

Single Point ◽

Single Point Mutation ◽

The Stability

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Specificity of repeat-induced point mutation (RIP) in Neurospora: sensitivity of non-Neurospora sequences, a natural diverged tandem duplication, and unique DNA adjacent to a duplicated region.

Genetics ◽

10.1093/genetics/127.4.711 ◽

1991 ◽

Vol 127 (4) ◽

pp. 711-717 ◽

Cited By ~ 1

Author(s):

E J Foss ◽

P W Garrett ◽

J A Kinsey ◽

E U Selker

Keyword(s):

Sequence Analysis ◽

Point Mutation ◽

Dna Sequence ◽

Dna Sequences ◽

Tandem Duplication ◽

Sexual Cycle ◽

Hygromycin B ◽

Plasmid Puc8 ◽

The Stability ◽

Repeat Induced Point Mutation

Abstract The process designated RIP (repeat-induced point mutation) alters duplicated DNA sequences in the sexual cycle of Neurospora crassa. We tested whether non-Neurospora sequences are susceptible to RIP, explored the basis for the observed immunity to this process of a diverged tandem duplication that probably arose by a natural duplication followed by RIP (the Neurospora zeta-eta region), and investigated whether RIP extends at all into unique sequences bordering a duplicated region. Bacterial sequences of the plasmid pUC8 and of a gene conferring resistance to hygromycin B were sensitive to RIP in N. crassa when repeated in the genome. When the entire 1.6-kb zeta-eta region was duplicated, it was susceptible to RIP, but was affected by it to a lesser extent than other duplications. Only three of 62 progeny from crosses harboring unlinked duplications of the region showed evidence of changes. We attribute the low level of alterations to depletion of mutable sites. The stability of the zeta-eta region in strains having single copies of the region suggests that the 14% divergence of the tandem elements is sufficient to prevent RIP. DNA sequence analysis of unduplicated pUC8 sequences adjacent to a duplication revealed that RIP continued at least 180 bp beyond the boundary of the duplication. Three mutations occurred in the 200-bp segment of bordering sequences examined.

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Open discussion

Symposium - International Astronomical Union ◽

10.1017/s0074180900029533 ◽

1982 ◽

Vol 99 ◽

pp. 605-613

Author(s):

P. S. Conti

Keyword(s):

Buenos Aires ◽

Large Mass ◽

List Type ◽

Common Phenomenon ◽

Open Discussion ◽

The Stability ◽

Ring Nebulae

Conti: One of the main conclusions of the Wolf-Rayet symposium in Buenos Aires was that Wolf-Rayet stars are evolutionary products of massive objects. Some questions:–Do hot helium-rich stars, that are not Wolf-Rayet stars, exist?–What about the stability of helium rich stars of large mass? We know a helium rich star of ∼40 MO. Has the stability something to do with the wind?–Ring nebulae and bubbles : this seems to be a much more common phenomenon than we thought of some years age.–What is the origin of the subtypes? This is important to find a possible matching of scenarios to subtypes.

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Symmetric Multistep Methods Revisited: II. Numerical Experiments

International Astronomical Union Colloquium ◽

10.1017/s0252921100031602 ◽

1999 ◽

Vol 173 ◽

pp. 309-314 ◽

Cited By ~ 3

Author(s):

T. Fukushima

Keyword(s):

Multistep Methods ◽

Computational Time ◽

Integration Time ◽

Implicit Methods ◽

Symmetric Methods ◽

Celestial Bodies ◽

The Stability ◽

Predictor Corrector ◽

Symmetric Multistep Methods ◽

Integration Errors

AbstractBy using the stability condition and general formulas developed by Fukushima (1998 = Paper I) we discovered that, just as in the case of the explicit symmetric multistep methods (Quinlan and Tremaine, 1990), when integrating orbital motions of celestial bodies, the implicit symmetric multistep methods used in the predictor-corrector manner lead to integration errors in position which grow linearly with the integration time if the stepsizes adopted are sufficiently small and if the number of corrections is sufficiently large, say two or three. We confirmed also that the symmetric methods (explicit or implicit) would produce the stepsize-dependent instabilities/resonances, which was discovered by A. Toomre in 1991 and confirmed by G.D. Quinlan for some high order explicit methods. Although the implicit methods require twice or more computational time for the same stepsize than the explicit symmetric ones do, they seem to be preferable since they reduce these undesirable features significantly.

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