- #Cellprofiler fiber length measurement generator
- #Cellprofiler fiber length measurement registration
These determine the importance of studying the replication fork dynamics and the factors that affect it. This suggests that the rapidly dividing cancer cells (and therefore having higher levels of replication stress) would be more susceptible to replication stress-inducing compounds and thus could be selectively destroyed ( Lecona and Fernandez-Capetillo 2014). On the other hand, excessive replication stress would not allow the completion of the S-phase, and the cell would die.
#Cellprofiler fiber length measurement generator
Damage caused by replication stress is considered to drive carcinogenesis, being the generator of genome instability required for the acquisition of the properties making a cell cancerous ( Gorgoulis, Vassiliou et al. Many intrinsic factors (e.g., specific DNA sequences and proteins that are tightly bound to DNA) as well as environmental factors (various chemical compounds) slow down or stop replication forks, causing a state known as “replication stress”.Ī consequence of replication stress is the accumulation of DNA damage ( Hills and Diffley 2014) as the unwound DNA at the sites of stopped replication complexes is more susceptible to being cut specific proteins target and break down stopped replication forks non-replicated portions of chromosomes are severed during mitosis. However, the movement of replication machines along the DNA molecule is not unimpeded. After initiation, replication forks then travel bidirectionally outwards from the origin ( Gambus, Jones et al. The unwinding of DNA at the origin by the CMG complex allows the replisome assembly, including the DNA synthesizing polymerases and the factors coordinating the various activities in the molecular machine that carries out DNA replication. 2006), which unwinds DNA double helix during replication. At the transition from G1 to S phases of the cell cycle, replication is triggered by the cyclin-dependent kinase (CDK) and Dbf4-dependent kinase (DDK), which activate the core replicative helicase – the MCM2-7 proteins, leading to the recruitment of additional factors and the formation of the active helicase – the CMG complex ( Gambus, Jones et al. The origins of replication are set during the G1 phase when pre-replication complexes are assembled on multiple sites along the genome, in a process known as licensing. The process is under strict control to ensure that every part of the genome is replicated only once in a cell cycle, and no parts remain unreplicated. In the S-phase of the cell cycle, replication in eukaryotes starts from multiple sites called origins of replication and runs in parallel. We hope that the application will facilitate both the study of DNA replication control and the effects of various compounds used in human activity on the process of DNA replication. It is freely available and does not require any paid platforms or services to be used. DNA size finder significantly simplified the analysis of the experimental data while increasing reliability by the standardized measurement of a greater number of DNA molecules. In this manuscript, we provide a detailed description of an algorithm and a lightweight Java application to automatically analyze single DNA molecule images we call “DNA size finder”. Recently, the interest in this kind of image analysis has grown rapidly. The major difficulty of the method is the labor-intensive analysis, which requires measuring the lengths of a large number of labeled fragments.
#Cellprofiler fiber length measurement registration
The method allows the registration of the activity of single replication complexes by measuring the length of the “trace” left by each of them. One of the most accurate methods for studying DNA replication is labeling newly synthesized DNA molecules with halogenated nucleotides, followed by immunofluorescence and microscopy detection, known as DNA fiber labeling. Disturbances of DNA replication have severe consequences for the affected cell, and current models link them to cancer development. Numerous control mechanisms ensure the exact course of the process during each cell division. The duplication of genetic information (DNA replication) is central to life.
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