Diana Huttner

Bio: Diana Huttner is an academic researcher from Technion – Israel Institute of Technology. The author has contributed to research in topics: Nanopore & DNA repair. The author has an hindex of 9, co-authored 10 publications receiving 487 citations. Previous affiliations of Diana Huttner include University of Copenhagen.

How unfinished business from S‐phase affects mitosis and beyond

Abstract: The eukaryotic cell cycle is conventionally viewed as comprising several discrete steps, each of which must be completed before the next one is initiated. However, emerging evidence suggests that incompletely replicated, or unresolved, chromosomes from S-phase can persist into mitosis, where they present a potential threat to the faithful segregation of sister chromatids. In this review, we provide an overview of the different classes of loci where this ‘unfinished S-phase business' can lead to a variety of cytogenetically distinct DNA structures throughout the various steps of mitosis. Furthermore, we discuss the potential ways in which cells might not only tolerate this inevitable aspect of chromosome biology, but also exploit it to assist in the maintenance of genome stability.

PICH promotes sister chromatid disjunction and co-operates with topoisomerase II in mitosis

Abstract: PICH is a SNF2 family DNA translocase that binds to ultra-fine DNA bridges (UFBs) in mitosis. Numerous roles for PICH have been proposed from protein depletion experiments, but a consensus has failed to emerge. Here, we report that deletion of PICH in avian cells causes chromosome structural abnormalities, and hypersensitivity to an inhibitor of Topoisomerase II (Topo II), ICRF-193. ICRF-193-treated PICH(-/-) cells undergo sister chromatid non-disjunction in anaphase, and frequently abort cytokinesis. PICH co-localizes with Topo IIα on UFBs and at the ribosomal DNA locus, and the timely resolution of both structures depends on the ATPase activity of PICH. Purified PICH protein strongly stimulates the catalytic activity of Topo II in vitro. Consistent with this, a human PICH(-/-) cell line exhibits chromosome instability and chromosome condensation and decatenation defects similar to those of ICRF-193-treated cells. We propose that PICH and Topo II cooperate to prevent chromosome missegregation events in mitosis.

Mechanistic insight into the interaction of BLM helicase with intra-strand G-quadruplex structures.

Abstract: Bloom syndrome is an autosomal recessive disorder caused by mutations in the RecQ family helicase BLM that is associated with growth retardation and predisposition to cancer. BLM helicase has a high specificity for non-canonical G-quadruplex (G4) DNA structures, which are formed by G-rich DNA strands and play an important role in the maintenance of genomic integrity. Here we used single-molecule FRET to define the mechanism of interaction of BLM helicase with intra-stranded G4 structures. We show that the activity of BLM is substrate dependent, and highly regulated by a short-strand DNA (ssDNA) segment that separates the G4 motif from double-stranded DNA. We demonstrate cooperativity between the RQC and HRDC domains of BLM during binding and unfolding of the G4 structure, where the RQC domain interaction with G4 is stabilized by HRDC binding to ssDNA. We present a model that proposes a unique role for G4 structures in modulating the activity of DNA processing enzymes.

Causes and consequences of replication stress.

Abstract: Replication stress is a complex phenomenon that has serious implications for genome stability, cell survival and human disease. Generation of aberrant replication fork structures containing single-stranded DNA activates the replication stress response, primarily mediated by the kinase ATR (ATM- and Rad3-related). Along with its downstream effectors, ATR stabilizes and helps to restart stalled replication forks, avoiding the generation of DNA damage and genome instability. Understanding this response may be key to diagnosing and treating human diseases caused by defective responses to replication stress.

What is a tutorial

Abstract: There are two kinds of tutorial articles: those that provide a primer on an established topic and those that let us in on the ground floor of something of emerging importance. The first type of tutorial can have a noted expert who has been gracious (and brave) enough to write a field guide about a particular topic. The other sort of tutorial typically involves researchers who have each been laboring on a topic for some years. Both sorts of tutorial articles are very much desired. But we, as an editorial board for both Systems and Transactions, know that there has been no logical place for them in the AESS until this series was started several years ago. With these tutorials, we hope to continue to give them a home, a welcome, and provide a service to our membership. We do not intend to publish tutorials on a regular basis, but we hope to deliver them once or twice per year. We need and welcome good, useful tutorial articles (both kinds) in relevant AESS areas. If you, the reader, can offer a topic of interest and an author to write about it, please contact us. Self-nominations are welcome, and even more ideal is a suggestion of an article that the editor(s) can solicit. All articles will be reviewed in detail. Criteria on which they will be judged include their clarity of presentation, relevance, and likely audience, and, of course, their correctness and scientific merit. As to the mathematical level, the articles in this issue are a good guide: in each case the author has striven to explain complicated topics in simple-well, tutorial-terms. There should be no (or very little) novel material: the home for archival science is the Transactions Magazine, and submissions that need to be properly peer reviewed would be rerouted there. Likewise, articles that are interesting and descriptive, but lack significant tutorial content, ought more properly be submitted to the Systems Magazine.

Methods and Applications

Abstract: As companies strive to develop artefacts intended for services instead of traditional sell-off, new challenges in the product development process arise to promote continuous improvement and increasing market profits. This creates a focus on product life-cycle components as companies then make life-cycle commitments, where they are responsible for the function availability during the extent of the life-cycle, i.e. functional products. One of these life-cycle components is manufacturing; therefore, companies search for new approaches of success during manufacturability evaluation already in engineering design. Efforts have been done to support early engineering design, as this phase sets constraints and opportunities for manufacturing. These efforts have turned into design for manufacturing methods and guidelines. A further step to improve the life-cycle focus during early engineering design is to reuse results and use experience from earlier projects. However, because results and experiences created during project work are often not documented for reuse, only remembered by some people, there is a need for design support. Knowledge engineering (KE) is a methodology for creating knowledge-based systems, e.g. systems that enable reuse of earlier results and make available both explicit and tacit corporate knowledge, enabling the automated generation and evaluation of new engineering design solutions during early product development. There are a variety of KE-approaches, such as knowledge-based engineering, case-based reasoning and programming, which have been used in research to develop design for manufacturing methods and applications. There are, however, opportunities for research where several approaches and their interdependencies, to create a transparent picture of how KE can be used to support engineering design, are investigated. The aim of the research presented in this thesis is to create new methods for design for manufacturing, by using several approaches of KE, and find the beneficial and less beneficial aspects of these methods in comparison to each other and earlier research. This thesis presents methods and applications for design for manufacturing using KE. KE has been employed in several ways, namely rule-based, rule-, programmingand finite element analysis (FEA)-based, and ruleand plan-based, which are tested and compared with each other. Results show that KE can be used to generate information about manufacturing in several ways. The rule-based way is suitable for supporting life-cycle commitments, as engineering design and manufacturing can be integrated with maintenance and performance predictions during early engineering design, though limited to the firing of production rules. The rule-, programmingand FEA-based way can be used to integrate computer-aided design tools and virtual manufacturing for non-linear stress and displacement analysis. This way may also bridge the gap between engineering designers and computational experts, even though this way requires a larger effort to program than the rule-based. The ruleand planbased way can enable design for manufacturing in two fashions – based on earlier manufacturing plans and based on rules. Because earlier manufacturing plans, together with programming algorithms, can handle knowledge that may be more intricate to capture as rules, as opposed to the time demanding routine work that is often automated by means of rules, several opportunities for designing for manufacturing exist.

Roles of eukaryotic topoisomerases in transcription, replication and genomic stability

Abstract: Topoisomerases introduce transient DNA breaks to relax supercoiled DNA, remove catenanes and enable chromosome segregation. Human cells encode six topoisomerases (TOP1, TOP1mt, TOP2α, TOP2β, TOP3α and TOP3β), which act on a broad range of DNA and RNA substrates at the nuclear and mitochondrial genomes. Their catalytic intermediates, the topoisomerase cleavage complexes (TOPcc), are therapeutic targets of various anticancer drugs. TOPcc can also form on damaged DNA during replication and transcription, and engage specific repair pathways, such as those mediated by tyrosyl-DNA phosphodiesterase 1 (TDP1) and TDP2 and by endonucleases (MRE11, XPF-ERCC1 and MUS81). Here, we review the roles of topoisomerases in mediating chromatin dynamics, transcription, replication, DNA damage repair and genomic stability, and discuss how deregulation of topoisomerases can cause neurodegenerative diseases, immune disorders and cancer.