Site-Specific Fluorescence Dynamics To Probe Polar Arrest by Fob1 in Replication Fork Barrier Sequences

ACS Omega
Anwesha BiswasRuchi Anand

Abstract

Fob1 protein plays an important role in aging and maintains genomic stability by avoiding clashes between the replication and transcription machinery. It facilitates polar arrest by binding to replication fork barrier (RFB) sites, present within the nontranscribed spacer region of the ribosomal DNA. Here, we investigate the mechanism of unidirectional arrest by creating multiple prosthetic forks within the RFB, with fluorescent adenine analogue 2-aminopurine incorporated site-specifically in both the "permissible" and "nonpermissible" directions. The motional dynamics of the RFB-Fob1 complexes analyzed by fluorescence lifetime and fluorescence anisotropy decay kinetics shows that Fob1 adopts a clamp-lock model of arrest and causes stronger perturbation with the bases in the double-stranded region of the nonpermissible-directed forks over those of the permissible directed ones, thereby creating a polar barrier. Corroborative thermal melting studies reveal a skewed distribution of GC content within the RFB sequence that potentially assists in Fob1-mediated arrest.

References

Jan 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·T D Petes
Nov 1, 1988·Molecular and Cellular Biology·M H Linskens, J A Huberman
Mar 26, 1984·Nucleic Acids Research·K G SkryabinA A Hadjiolov
Jan 15, 1998·Cell·D A Sinclair, L Guarente
Mar 21, 1998·Nucleic Acids Research·B HolzE Weinhold
Mar 16, 2002·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Katsuki Johzuka, Takashi Horiuchi
Apr 23, 2002·Protein Science : a Publication of the Protein Society·Mensur Dlakić
Jun 5, 2003·Genes & Development·Yasushi TakeuchiTakehiko Kobayashi
Apr 3, 2007·EMBO Reports·Karim Labib, Ben Hodgson
May 2, 2007·The Journal of Physical Chemistry. B·T RamreddyG Krishnamoorthy
Oct 9, 2007·Journal of Molecular Biology·Nabanita NagG Krishnamoorthy
Feb 28, 2009·Proceedings of the National Academy of Sciences of the United States of America·Davis JosePeter H von Hippel
May 8, 2009·The Journal of Physical Chemistry. B·T RamreddyB J Rao
Jun 24, 2010·The Journal of Physical Chemistry. B·Teena GoelGuruswamy Krishnamoorthy
Aug 22, 2012·Proceedings of the National Academy of Sciences of the United States of America·Davis JosePeter H von Hippel

❮ Previous
Next ❯

Citations


❮ Previous
Next ❯

Methods Mentioned

BETA
X-ray
electrophoretic mobility
Fluorescence
surface plasmon resonance
Assay
Fluorescence Decay

Software Mentioned

BLAST
ImageQuant

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.