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Structure-function analysis of the SaPIbov1 replication origin in Staphylococcus aureus.

Carles Ubeda¹, María Ángeles Tormo-Más, José R Penadés

¹Molecular Pathogenesis Program, Skirball Institute, New York University School of Medicine, 540 First Ave., New York, NY 10016, USA.

|January 28, 2012

Summary

This summary is machine-generated.

Bacterial pathogenicity islands called SaPIs replicate autonomously using a phage-like origin. Their initiator protein (Rep) binds to specific sites, enabling DNA replication initiation.

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Bacterial pathogenicity islands (SaPIs) are unique mobile genetic elements.
Unlike other pathogenicity islands, SaPIs can replicate autonomously.

Purpose of the Study:

To investigate the mechanism of autonomous replication of SaPIs.
To understand the role of the SaPI-specific initiator protein (Rep) and its binding sites in replication initiation.

Main Methods:

Deletion analysis of the SaPI origin of replication.
Gel mobility assays to study Rep protein binding to iterons.
Investigating replication initiation on linear DNA.

Main Results:

The SaPI replicon contains two sets of iterons flanking an AT-rich region.
Rep protein binds to single iterons, and binding to both sites is necessary for replication initiation.
The entire origin sequence is required for Rep-driven melting and initiation.
SaPI replication can be initiated on linear DNA.

Conclusions:

SaPI replication initiation requires specific binding of the Rep protein to both flanking iterons.
Bilateral Rep binding likely constrains the DNA, facilitating melting of the AT-rich region.
The findings elucidate a novel mechanism for autonomous replication of mobile genetic elements.