Initiation of DNA replication at a nuclear matrix-attached chromatin fraction

2005 ◽  
Vol 203 (1) ◽  
pp. 71-77 ◽  
Author(s):  
Ilian Radichev ◽  
Anastassia Parashkevova ◽  
Boyka Anachkova
1998 ◽  
Vol 18 (7) ◽  
pp. 4165-4176 ◽  
Author(s):  
Lin-Feng Chen ◽  
Kosei Ito ◽  
Yota Murakami ◽  
Yoshiaki Ito

ABSTRACT The nuclear matrix is thought to play an important role in the DNA replication of eukaryotic cells, although direct evidence for such a role is still lacking. A nuclear matrix-associated transcription factor, polyomavirus (Py) enhancer binding protein 2αB1 (PEBP2αB1) (AML1/Cbfa2), was found to stimulate Py replication through its cognate binding site. The minimal replication activation domain (RAD) was identified between amino acid (aa) 302 and aa 371 by using a fusion protein containing the GAL4 DNA binding domain (GAL4-RAD). In addition, the region showed affinity for the nuclear matrix and, on the basis of competition studies, binding activity for one or more proteins involved in the initiation of Py DNA replication. A leukemogenic chimeric protein, AML1/ETO(MTG8), which does not contain this region of PEBP2αB1/AML1, was also localized in the nuclear matrix fraction and competed for nuclear matrix association with PEBP2αB1 and GAL4-RAD. Moreover, AML1/ETO inhibited Py DNA replication stimulated by PEBP2αB1 and GAL4-RAD. The inhibition was specific for replication mediated by PEBP2αB1 and GAL4-RAD, and proportional to the degree of loss of these activators from the nuclear matrix, suggesting a requirement for nuclear matrix targeting in the stimulation of Py DNA replication by RAD. These results are the first to suggest a molecular link between the initiation of DNA replication and the nuclear matrix compartment.


2019 ◽  
Vol 16 (3) ◽  
pp. 272-277 ◽  
Author(s):  
Rasmus N. Klitgaard ◽  
Anders Løbner-Olesen

Background:One of many strategies to overcome antibiotic resistance is the discovery of compounds targeting cellular processes, which have not yet been exploited.Materials and Methods:Using various genetic tools, we constructed a novel high throughput, cellbased, fluorescence screen for inhibitors of chromosome replication initiation in bacteria.Results:The screen was validated by expression of an intra-cellular cyclic peptide interfering with the initiator protein DnaA and by over-expression of the negative initiation regulator SeqA. We also demonstrated that neither tetracycline nor ciprofloxacin triggers a false positive result. Finally, 400 extracts isolated mainly from filamentous actinomycetes were subjected to the screen.Conclusion:We concluded that the presented screen is applicable for identifying putative inhibitors of DNA replication initiation in a high throughput setup.


2002 ◽  
Vol 277 (34) ◽  
pp. 30824-30831 ◽  
Author(s):  
Ming Lei ◽  
Irene H. Cheng ◽  
Louis A. Roberts ◽  
Michael A. McAlear ◽  
Bik K. Tye

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