Interaction of the H4 autonomously replicating sequence core consensus sequence and its 3'-flanking domain

1989 ◽  
Vol 9 (12) ◽  
pp. 5464-5472
Author(s):  
S G Holmes ◽  
M M Smith
Keyword(s):  
Consensus Sequence ◽  
Point Mutations ◽  
Relative Orientation ◽  
Autonomously Replicating Sequence ◽  
The Core ◽  
Multiple Copies ◽  
Minimal Sequences ◽  
Necessary And Sufficient ◽  
The Relationship

Yeast autonomously replicating sequence (ARS) elements are composed of a conserved 11-base-pair (bp) core consensus sequence and a less well defined 3'-flanking region. We have investigated the relationship between the H4 ARS core consensus sequence and its 3'-flanking domain. The minimal sequences necessary and sufficient for function were determined by combining external 3' and 5' deletions to produce a nested set of ARS fragments. Sequences 5' of the core consensus were dispensable for function, but at least 66 bp of 3'-flanking domain DNA was required for full ARS function. The importance of the relative orientation of the core consensus element with respect to the 3'-flanking domain was tested by precisely inverting 14 bp of DNA including the core consensus sequence by oligonucleotide mutagenesis. This core inversion mutant was defective for all ARS function, showing that a fixed relative orientation of the core consensus and 3'-flanking domain is required for function. The 3'-flanking domain of the minimal functional H4 ARS fragment contains three sequences with a 9-of-11-bp match to the core consensus. The role of these near-match sequences was tested by directed mutagenesis. When all near-match sequences with an 8-of-11-bp match or better were simultaneously disrupted by point mutations, the resulting ARS construct retained full replication function. Therefore, multiple copies of a sequence closely related to the core consensus element are not required for H4 ARS function.

scholarly journals Interaction of the H4 autonomously replicating sequence core consensus sequence and its 3'-flanking domain.

1989 ◽  
Vol 9 (12) ◽  
pp. 5464-5472 ◽  
Author(s):  
S G Holmes ◽  
M M Smith
Keyword(s):  
Consensus Sequence ◽  
Point Mutations ◽  
Relative Orientation ◽  
Autonomously Replicating Sequence ◽  
The Core ◽  
Multiple Copies ◽  
Minimal Sequences ◽  
Necessary And Sufficient ◽  
The Relationship

Yeast autonomously replicating sequence (ARS) elements are composed of a conserved 11-base-pair (bp) core consensus sequence and a less well defined 3'-flanking region. We have investigated the relationship between the H4 ARS core consensus sequence and its 3'-flanking domain. The minimal sequences necessary and sufficient for function were determined by combining external 3' and 5' deletions to produce a nested set of ARS fragments. Sequences 5' of the core consensus were dispensable for function, but at least 66 bp of 3'-flanking domain DNA was required for full ARS function. The importance of the relative orientation of the core consensus element with respect to the 3'-flanking domain was tested by precisely inverting 14 bp of DNA including the core consensus sequence by oligonucleotide mutagenesis. This core inversion mutant was defective for all ARS function, showing that a fixed relative orientation of the core consensus and 3'-flanking domain is required for function. The 3'-flanking domain of the minimal functional H4 ARS fragment contains three sequences with a 9-of-11-bp match to the core consensus. The role of these near-match sequences was tested by directed mutagenesis. When all near-match sequences with an 8-of-11-bp match or better were simultaneously disrupted by point mutations, the resulting ARS construct retained full replication function. Therefore, multiple copies of a sequence closely related to the core consensus element are not required for H4 ARS function.

scholarly journals Mutational analysis of the consensus sequence of a replication origin from yeast chromosome III.

1990 ◽  
Vol 10 (8) ◽  
pp. 3917-3925 ◽  
Author(s):  
J V Van Houten ◽  
C S Newlon
Keyword(s):  
Base Pair ◽  
High Frequency ◽  
Mutational Analysis ◽  
Plasmid Stability ◽  
Consensus Sequence ◽  
Point Mutations ◽  
Autonomously Replicating Sequence ◽  
Frequency Transformation ◽  
The Core ◽  
High Frequency Transformation

Yeast autonomously replicating sequence (ARS) elements contain an 11-base-pair core consensus sequence (5'-[A/T]TTTAT[A/G]TTT[A/T]-3') that is required for function. The contribution of each position within this sequence to ARS activity was tested by creating all possible single-base mutations within the core consensus sequence of ARS307 (formerly called the C2G1 ARS) and testing their effects on high-frequency transformation and on plasmid stability. Of the 33 mutations, 22 abolished ARS function as measured by high-frequency transformation, 7 caused more than twofold reductions in plasmid stability, and 4 had no effect on plasmid stability. Mutations that reduced or abolished ARS activity occurred at each position in the consensus sequence, demonstrating that each position of this sequence contributes to ARS function. Of the four mutations that had no effect on ARS activity, three created alternative perfect matches to the core consensus sequence, demonstrating that the alternate bases allowed by the consensus sequence are, indeed, interchangeable. In addition, a change from T to C at position 6 did not perturb wild-type efficiency. To test whether the essential region extends beyond the 11-base-pair consensus sequence, the effects on plasmid stability of point mutations one base 3' to the T-rich strand of the core consensus sequence (position 12) and deletion mutations that altered bases 5' to the T-rich strand of the core consensus sequence were examined. An A at position 12 or the removal of three T residues 5' to the core consensus sequence severely diminished ARS efficiency, showing that the region required for full ARS efficiency extends beyond the core consensus sequence in both directions.

Mutational analysis of the consensus sequence of a replication origin from yeast chromosome III

1990 ◽  
Vol 10 (8) ◽  
pp. 3917-3925
Author(s):  
J V Van Houten ◽  
C S Newlon
Keyword(s):  
Base Pair ◽  
High Frequency ◽  
Mutational Analysis ◽  
Plasmid Stability ◽  
Consensus Sequence ◽  
Point Mutations ◽  
Autonomously Replicating Sequence ◽  
Frequency Transformation ◽  
The Core ◽  
High Frequency Transformation

Yeast autonomously replicating sequence (ARS) elements contain an 11-base-pair core consensus sequence (5'-[A/T]TTTAT[A/G]TTT[A/T]-3') that is required for function. The contribution of each position within this sequence to ARS activity was tested by creating all possible single-base mutations within the core consensus sequence of ARS307 (formerly called the C2G1 ARS) and testing their effects on high-frequency transformation and on plasmid stability. Of the 33 mutations, 22 abolished ARS function as measured by high-frequency transformation, 7 caused more than twofold reductions in plasmid stability, and 4 had no effect on plasmid stability. Mutations that reduced or abolished ARS activity occurred at each position in the consensus sequence, demonstrating that each position of this sequence contributes to ARS function. Of the four mutations that had no effect on ARS activity, three created alternative perfect matches to the core consensus sequence, demonstrating that the alternate bases allowed by the consensus sequence are, indeed, interchangeable. In addition, a change from T to C at position 6 did not perturb wild-type efficiency. To test whether the essential region extends beyond the 11-base-pair consensus sequence, the effects on plasmid stability of point mutations one base 3' to the T-rich strand of the core consensus sequence (position 12) and deletion mutations that altered bases 5' to the T-rich strand of the core consensus sequence were examined. An A at position 12 or the removal of three T residues 5' to the core consensus sequence severely diminished ARS efficiency, showing that the region required for full ARS efficiency extends beyond the core consensus sequence in both directions.

scholarly journals The Content and the Role of Trust Management Service in the Sector of Individual Accommodation Facilities in the Market of Hotel Services

2019 ◽  
Vol 29 (2) ◽  
pp. 305-313
Author(s):  
Nina Danilenko ◽  
Oxana Suranova
Keyword(s):  
Trust Management ◽  
Commercial Real Estate ◽  
Management Service ◽  
General Scientific ◽  
The Individual ◽  
Necessary And Sufficient ◽  
The Relationship ◽  
Individual Placement ◽  
Management Services

The article defines the content and the role of trust management service in the sector of individual accommodation facilities. The object of the trust management service is residential property owned by households. The relationship between households and management companies in the service is assessed as a factor in ensuring socio-economic efficiency in the individual placement sector. During the study, the author has used a complex of general scientific research methods (analysis, synthesis, the comparative analytical method, the analogy method). Based on the analysis of individual means of accommodation as residential and commercial real estate, the article proves the need for trust management services and clarifies its content. It determines the list of necessary and sufficient indicators for evaluating the effectiveness of trust management services in the sector of individual accommodation facilities. The development of trust management services in the sector of individual accommodation will facilitate the provision of comfortable and safe services in the market of hotel services.

scholarly journals The Saccharomyces cerevisiae SPT8 gene encodes a very acidic protein that is functionally related to SPT3 and TATA-binding protein.

Genetics ◽  
1994 ◽  
Vol 137 (3) ◽  
pp. 647-657 ◽  
Author(s):  
D M Eisenmann ◽  
C Chapon ◽  
S M Roberts ◽  
C Dollard ◽  
F Winston
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Binding Protein ◽  
Tata Binding Protein ◽  
Sequence Motif ◽  
Heterotrimeric G Proteins ◽  
Eukaryotic Proteins ◽  
Multiple Copies ◽  
The Relationship ◽  
Insertion Mutations

Abstract Mutations in the Saccharomyces cerevisiae SPT8 gene were previously isolated as suppressors of retrotransposon insertion mutations in the 5' regions of the HIS4 and LYS2 genes. Mutations in SPT8 confer phenotypes similar to those caused by particular mutations in SPT15, which encodes the TATA-binding protein (TBP). These phenotypes are also similar to those caused by mutations in the SPT3 gene, which encodes a protein that directly interacts with TBP. We have now cloned and sequenced the SPT8 gene and have shown that it encodes a predicted protein of 602 amino acids with an extremely acidic amino terminus. In addition, the predicted SPT8 amino acid sequence contains one copy of a sequence motif found in multiple copies in a number of other eukaryotic proteins, including the beta subunit of heterotrimeric G proteins. To investigate further the relationship between SPT8, SPT3 and TBP, we have analyzed the effect of an spt8 null mutation in combination with different spt3 and spt15 mutations. This genetic analysis has shown that an spt8 deletion mutation is suppressed by particular spt3 alleles. Taken together with previous results, these data suggest that the SPT8 protein is required, directly or indirectly, for TBP function at particular promoters and that the role of SPT8 may be to promote a functional interaction between SPT3 and TBP.

scholarly journals Mitos dan Budaya

Humaniora ◽  
2015 ◽  
Vol 6 (2) ◽  
pp. 190 ◽  
Author(s):  
Mia Angeline
Keyword(s):  
Human Activity ◽  
Daily Activities ◽  
Modern Culture ◽  
The Core ◽  
The World ◽  
Almost All ◽  
The Relationship ◽  
Universal Themes ◽  
The Universe

This article describes the role of myth and some universal themes of myth, such as the creation of the world, a huge flood, death, and the end of the world. Almost all the world's myths concern this universal themes, as seen from the similarity of some of the myths followed by many cultures in the world. These myths have primary functions to human’s behavior and attitude because people keep telling almost the same myths to their predecessors. The goals for this research are (1) knowing the functions of myths with famous themes from various culture and (2) knowing the background and relationship between myths and modern culture. The result describes the relationship between the cultural myths, where the core of the story is the truth of humanity. In addition, myth acts as a template to organize their daily activities as well as human activity, but it also serves to introduce human to a greater power in the universe. The values in each story will be interpreted as rules and customs that must be met, and this has resulted in the emergence of a culture passed down from generation to generation. 

Back to Practices and Narratives

2022 ◽  
pp. 265-279
Author(s):  
Elisabetta Risi ◽  
Riccardo Pronzato
Keyword(s):  
Critical Pedagogy ◽  
Everyday Life ◽  
Daily Activities ◽  
Digital Platforms ◽  
The Core ◽  
Research Fields ◽  
Auto Ethnography ◽  
Core Idea ◽  
The Relationship

The role of digital platforms in everyday life is a concern within different research fields; therefore, several authors have supported the need to investigate them and their underlying meshing of human and computational logic. In this chapter, the authors present a methodological proposal according to which auto-ethnographic diaries can be fruitfully employed to examine the relationship between individuals and algorithmic platforms. By drawing on a critical pedagogy approach, they consider auto-ethnography both as a practice of access to algorithmic logics through rich first-hand data regarding everyday usage practices as a response to datafication. The core idea behind this narrative method is to use inductive self-reflexive methodological tools to help individuals critically reflect on their daily activities, thereby making their consumption of algorithmic contents more aware and allowing researchers to collect in-depth reports about their use of digital platforms and the following processes of subjectification.

scholarly journals cis-acting components in the replication origin from ribosomal DNA of Saccharomyces cerevisiae.

1993 ◽  
Vol 13 (9) ◽  
pp. 5360-5369 ◽  
Author(s):  
C A Miller ◽  
D Kowalski
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nucleotide Sequence ◽  
Ribosomal Dna ◽  
Consensus Sequence ◽  
Perfect Match ◽  
Point Mutations ◽  
Dna Unwinding ◽  
Mung Bean Nuclease ◽  
Autonomously Replicating Sequence ◽  
Cis Acting

The ribosomal DNA (rDNA) repeats of Saccharomyces cerevisiae contain an autonomously replicating sequence (ARS) that colocalizes with a chromosomal origin of replication. We show that a minimal sequence necessary for full ARS function corresponds to a 107-bp rDNA fragment which contains three 10-of-11-bp matches to the ARS consensus sequence. Point mutations in only one of the 10-of-11-bp matches, GTTTAT GTTTT, inactivate the rDNA ARS, indicating that this consensus sequence is essential. A perfect match to a revised ARS consensus is present but not essential. Sequences up to 9 bp 5' from the essential consensus are dispensable. A broad DNA region directly 3' to the essential consensus is required and is easily unwound as indicated by: (i) hypersensitivity to nicking of an approximately 100-bp region by mung bean nuclease in a negatively supercoiled plasmid and (ii) helical instability determined by thermodynamic analysis of the nucleotide sequence. A correlation between DNA helical instability and replication efficiency of wild-type and mutated ribosomal ARS derivatives suggests that a broad region 3' to the essential ARS consensus functions as a DNA unwinding element. Certain point mutations that do not stabilize the DNA helix in the 3' region but reduce ARS efficiency reveal an element distinct from, but overlapping, the DNA unwinding element. The nucleotide sequence of the functionally important constituents in the ARS appears to be conserved among the rDNA repeats in the chromosome.

International Organizations and their Members

2004 ◽  
Vol 1 (1) ◽  
pp. 139-161 ◽  
Author(s):  
Niels Blokker
Keyword(s):  
International Organizations ◽  
Legal Status ◽  
English Word ◽  
The Body ◽  
International Organization ◽  
The Core ◽  
The Law ◽  
Core Questions ◽  
The Relationship

AbstractThe law of international organizations is a field of study full of themes with variations. With respect to a number of key chapters in this field, such as the legal status of international organizations, their institutional structure, powers, decision-making and decisions, sanctions, fundamental principles exist or are being developed. These principles or themes have a large number of variations in the rules and practice of individual organizations. Like in music, the variations in themselves may be interesting as such, but they can only be fully appreciated by not losing sight of the theme. Like in music, the themes in themselves may be interesting as such, but they can only be fully appreciated by also listening carefully to the multicoloured variations. This contribution is devoted to a central theme of the law of international organizations: the relationship between an international organization and its members. Already in the early days of existence of international organizations, this topic has given rise to numerous questions. Many of these have now been answered, but sometimes resurface in contemporary variations. In addition there are new questions. It is useful to first explore briefly the meaning of the word "members". The English word "member" and the French "membre" are both derived from the Latin word "membrum", which means "part of the body". This meaning is important because it indicates clearly that members are part of a whole – a fact which takes us straight to one of the core questions addressed in this article. Members of an international organization are not just members, after all – like the members of a bridge club or a gardening club. In most cases, the members of international organizations are states. How then can members of an international organization be members of a whole when in most cases those members are sovereign states, even if the concept of sovereignty of states is no longer as absolute at it used to be? This is one of the major questions within the field of the law of international organizations. In order to examine this question, this contribution is divided into two parts. Section I will concentrate on the role of members vis-à-vis international organizations. In Section II the focus will shift to the whole of which the members are part; this Section will briefly look at some of the questions involved in giving a certain amount of autonomy to this whole.

scholarly journals Whither neoliberal penality? The past, present and future of imprisonment in the US

2018 ◽  
Vol 21 (2) ◽  
pp. 187-206 ◽  
Author(s):  
Sappho Xenakis ◽  
Leonidas K Cheliotis
Keyword(s):  
State Regulation ◽  
Donald Trump ◽  
Government Institutions ◽  
The Core ◽  
The Past ◽  
The Us ◽  
Economic Developments ◽  
The Relationship ◽  
Considerable Doubt

Debates about the trajectory of prison rates in the US, on one hand, and about the prospects of the neoliberal international order, on the other hand, suggest the time is ripe for a reappraisal of penological scholarship on the relationship between neoliberalism and imprisonment. With the aim of responding to this challenge, this article considers the relevance of the so-called ‘neoliberal penality thesis’ as a framework through which to interpret recent and ongoing developments in US imprisonment. We first set out the core propositions of the thesis and engage with a range of critiques it has attracted regarding the role of crime and government institutions, the evolution and functions of state regulation and welfare provision, and reliance on imprisonment as an indicator of state punitiveness. We then outline the principal arguments that have arisen about the direction of contemporary prison trends in the US, including since Donald Trump was elected to the presidency and took office, and proceed to distil their commonly opaque treatment of the intersections between neoliberalism and imprisonment, also clarifying their respective implications for the neoliberal penality thesis in light of the main critiques levelled previously against it. In so doing, we go beyond the penological field to take into account concerns about the vitality of neoliberalism itself. We conclude that international politico-economic developments have cast considerable doubt over the pertinence of neoliberalism as an organising concept for analysis of emergent penal currents.

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