Active Cells, Biochemistry & Physiology Infection & Immunity

Mechanism of pilus biogenesis by bacterial conjugative transfer systems

In plain English

AI plain-English summary

Bacteria use a hollow protein filament called a pilus to physically connect with neighbouring cells and funnel DNA—including antibiotic resistance genes—directly into them. This matters because the rapid spread of resistance genes among bacterial pathogens is driven largely by this process, called conjugation. Without a pilus, no transfer can occur. The molecular machine that builds the pilus is a complex of ten proteins embedded in the bacterial membrane. The researchers have already solved the atomic structure of this entire machine using cryo-electron microscopy, and now they want to work out exactly how it assembles the pilus filament step by step. This is fundamental science. Understanding the precise mechanism of pilus construction will not immediately produce a new drug or treatment. But it will reveal specific molecular targets—points in the assembly process that could be blocked by future inhibitors. If researchers can later design molecules that jam pilus formation, they could stop the horizontal spread of antibiotic resistance genes between bacteria, slowing the crisis without directly killing the bacteria themselves. Similar structural work on bacterial secretion systems has already led to candidate antivirulence compounds.

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The rise of antibiotic resistance is one of the major threats to public health. Antibiotics have been the standard of care for bacterial infections for decades. Since their discovery, hundreds of millions of humans have successfully been treated by these "wonder" drugs. Unfortunately, the rapid spread of antibiotic resistance genes among bacterial pathogens threatens to undo the remarkable progress made over the years to combat bacterial infections. Conjugation (or conjugative transfer) is a widespread biological process whereby genes are exchanged horizontally and unidirectionally from a bacterial donor cell to a bacterial recipient cell. Because conjugation is the process via which antibiotic resistance genes propagate among bacterial pathogens, it is in large part responsible for the antibiotic resistance crisis that humanity is presently experiencing. Crucial to conjugative transfer is the prior formation of a pilus, a long hollow filament produced by the donor cell. The pilus is made of 1000s of subunits of a protein called VirB2. The pilus serves not only to make contact with the recipient cell but also as a conduit for the transfer of DNAs from donor to recipient cells. A pilus needs to be generated before DNA transfer can begin. Pilus biogenesis and DNA transfer are mediated by a very large molecular machine called "conjugative transfer system" which is embedded in the membranes of the donor cell. Although 12 proteins, named VirB1-11 and VirD4, are necessary to carry out pilus biogenesis and DNA transfer, only the VirB2-11 proteins are required for pilus biogenesis. Recently, we have determined the cryo-EM atomic resolution structure of a large complex containing all the 10 proteins involved in pilus biogenesis and, as a result, we were able to propose a plausible mechanism for pilus biogenesis. Here we propose to build on this recent work and elucidate the details of the mechanism of pilus biogenesis by these complex systems. Mechanistic and structural details will lead to a better understanding of how antibiotics resistance genes spread among bacterial populations and therefore will provide the means to block it.

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Researchers

Gabriel Waksman (Principal Investigator)Konstantinos Thalassinos (Co-Investigator)

Related Research

Grants with similar aims, by meaning.

Molecular Mechanism Of DNA Processing For Antibiotic Resistance Gene Transfer In Bacteria
Structural and molecular investigations of membrane-embedded pilus assembly nanomachines in Gram-negative bacterial pathogens
Understanding the molecular mechanism of antibiotic resistance gene transfer in bacteria
Unravelling the molecular basis of subunit specificity in bacterial pilus assembly mechanisms
The bacterial type IV pilus machinery as a DNA translocator

Original classification

Research Grant

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