Study reveals new survival strategy for bacteria exposed to antibiotics

Researchers have uncovered a new way that some bacteria survive when under siege by antibiotics.

This survival mechanism is fundamentally different from other, known bacterial strategies. Understanding it may be useful for designing drugs that target hard-to-treat bacterial strains, such as drug-resistant tuberculosis, an increasingly urgent public health problem. The study is based on Mycobacterium smegmatis, a cousin of the microbe that causes TB, and its response to the TB drug isoniazid.

The research, by Yuichi Wakamoto of the University of Tokyo and Neeraj Dhar of the Swiss Federal Institute of Technology in Lausanne and colleagues, appears in the 4 January issue of Science. The journal is published by AAAS, the nonprofit, international science society.

Researchers observed as early as 1944 that antibiotics are less effective against cell populations that aren't proliferating. More recently, experiments have shown that some bacteria survive exposure to antibiotics thanks to a population of non-dividing "persister cells" that are present in the population even before the antibiotic treatment begins.

"This concept has been widely accepted as a general explanation for bacterial persistence despite very limited experimental support," said Wakamoto.

Wakamoto and colleagues now report that non-dividing persister cells are not responsible for the survival of M. smegmatis exposed to isoniazid. In fact, cell survival is not related to growth rate at all. Instead, random pulses of a bacterial enzyme called KatG make it possible for some cells to survive antibiotic treatment.

"Our Science paper provides clear experimental proof that other mechanisms of persistence also exist," said Dhar. "Our findings necessitate the re-examination of the mechanisms of persistence at the single-cell level in other bacteria, including Mycobacterium tuberculosis, which causes TB in humans."

The researchers studied single M. smegmatis cells in microfluidic cultures, treated with isoniazid. This drug is a "pro-drug" that does not become active until it is administered and interacts with certain compounds in the cell. In the case of M. smegmatis, it's KatG that activates isoniazid.

Individual cells' fates were not correlated with their growth rates but rather with their production of KatG. Each cell produced KatG in random pulses that determined the cell's chances of survival.

The researchers conclude that in certain cells, there were periods in between pulses when enzyme conversion of the pro-drug was barely possible. Thus a few cells probably avoided being killed by the activated antibiotic.

"At present we can only speculate as to whether the same or similar mechanisms exist in other bacterial species, although we think this is likely," said Wakamoto.

This research was supported by the Charles H. Revson Foundation, the Heiser Program for Research in Leprosy and Tuberculosis of the New York Community Trust, the Harvey L. Karp Discovery Fellowship and the JST PRESTO Program, the Bill and Melinda Gates Foundation, the National Institutes of Health, and the Swiss National Science Foundation.

The American Association for the Advancement of Science (AAAS) is the world's largest general scientific society and publisher of the journal Science (www.sciencemag.org) as well as Science Translational Medicine (www.sciencetranslationalmedicine.org) and Science Signaling (www.sciencesignaling.org). AAAS was founded in 1848 and includes some 261 affiliated societies and academies of science, serving 10 million individuals. Science has the largest paid circulation of any peer-reviewed general science journal in the world, with an estimated total readership of 1 million. The non-profit AAAS (www.aaas.org) is open to all and fulfills its mission to "advance science and serve society" through initiatives in science policy, international programs, science education, public engagement, and more. For the latest research news, log onto EurekAlert!, www.eurekalert.org, the premier science-news Web site, a service of AAAS. See www.aaas.org.