Ancient Viral DNA: From Genome Invaders to Gene Whisperers

Over the past decade, scientists discovered how bacteria defend themselves against viruses using CRISPR, a system that has revolutionized gene editing. But what if bacteria and viruses have been repurposing their own genetic tools for entirely different functions? A new study published in Nature by Columbia postdocs Tanner Wiegand, Egill Richard and Chance Meers uncovers a surprising twist in this evolutionary story: ancient genes, once used by viruses and mobile DNA elements called transposons, have evolved into RNA-guided transcriptional regulators – natural gene regulators that operate much like the immune system CRISPR, but without cutting DNA.

Bacteria are constantly in an evolutionary arms race with viruses. The mobile DNA pieces transposons have played a major role in bacterial evolution by jumping between locations in the genome and spreading genes. One of these genes, TnpB, was long thought to function only as a simple DNA-cutting enzyme, helping transposons move around. However, scientists have now discovered that some TnpB-derived proteins have lost their cutting ability and instead evolved into RNA-guided transcription factors – proteins that turn genes on or off by blocking access to DNA.

Wiegand and colleagues named this new class of proteins TldRs (TnpB-like nuclease-dead repressors). Unlike CRISPR proteins, which evolved from transposon-related genes to protect bacteria from viruses, TldRs appear to have been repurposed independently for gene regulation. These proteins rely on small RNA molecules to guide them to specific sequences in bacterial genomes, where they bind and shut down target genes by preventing transcription.

One of the most fascinating discoveries in the study is how viruses have co-opted TldRs to manipulate their bacterial hosts. The researchers found that some bacteriophages – viruses that infect bacteria – carry TldR genes along with a bacterial gene called fliC, which encodes flagellin, the protein that makes up the bacterial flagellum (a whip-like structure used for movement).

Normally, bacteria express their own version of fliC to build flagella, which are crucial for swimming and sensing the environment. However, in bacteria infected with certain bacteriophages, the viral version of fliC (fliCᴾ) replaces the host’s version. The study shows that TldRs, guided by their RNA molecules, specifically silence the bacterial fliC gene while allowing the viral version to be expressed. This means that the bacterium’s flagella are partially “rewired” to contain viral proteins instead of its own (Figure 1).

Why would a virus want to do this? In their study, Wiegand et al. propose a few possibilities:

  • Evading the immune system – Many bacterial flagellins are recognized by immune cells, and replacing them with a different version could help bacteria (and their resident viruses) go unnoticed.
  • Avoiding other viral infections – Some bacteriophages recognize flagella as entry points to infect bacteria. By altering flagellar proteins, the virus controlling the bacterium might block access to competing viruses.
  • Affecting bacterial motility – Flagella are primarily involved in motility. Changing their structure could alter how bacteria swim in various environments.

Figure 1: Schematic representation of the repression of the bacterial fliC gene by the RNA-guided viral-encoded TldR and consequent expression of the viral version of the flagellum gene, fliCp. Figure from the original paper. 

One of the most exciting aspects of this discovery of this new class of gene regulators is their potential for biotechnology tools development. CRISPR-based gene editing relies on programmable RNA-guided proteins, and TldRs appear to work on the same principle – but without the need to cut DNA. Instead, they act as natural gene “dimmer switches”, fine-tuning expression levels in a targeted way. Because TldRs are much smaller than CRISPR-Cas proteins, they could be useful for future genetic engineering applications where space is limited, such as in gene therapy or synthetic biology. Unlike traditional CRISPR editing, which involves cutting DNA (which can lead to unwanted mutations), TldRs could provide a more precise way to repress or regulate genes without permanent genome changes.

This study is a striking reminder of evolution’s endless innovations and how nature continuously repurposes molecular tools over evolutionary time. The same transposon-derived genes that gave rise to CRISPR have now been independently adapted to regulate gene expression in bacteria. It also underscores how much we still have to learn about the diversity of RNA-guided systems beyond CRISPR, which could lead to new technologies inspired by nature’s own innovations.

With further research, TldRs might become a new class of programmable genetic tools, opening new doors in medicine, synthetic biology, and biotechnology. As we continue to explore these hidden layers of microbial evolution, who knows what other surprises nature has in store?

Reviewed by: Temistocles Molinar, Saheli Chowdhury

A Germ of Understanding in the Gut Microbiome

Bacteria haven’t had much luck with the press.  Since the advent of germ theory in the 1800s, pathogenic bacteria – the disease-causing “bad apples” of the bacterial barrel – have hogged the spotlight in both science and popular imagination. This trend has started shifting over the last decade, as researchers have turned their attention toward the human microbiome: the trillions of bacteria, archaea, and other microbes which reside inside the human body (primarily the gastrointestinal tract) and are now known to contribute to host immunity, metabolism, and even behavior.

But how? By what mechanisms do these microscopic guests communicate with their host to exert such profound effects?  In contrast to well-characterized host-pathogen interactions, interactions between hosts and commensal (i.e., “friendly”) microbes have remained largely unexplored. In a recent collaborative study, Mark Ladinsky of the California Institute of Technology and Dr. Leandro Araujo of Columbia University sought to change that.

Ladinsky and Araujo focused their investigation on one class of microbes in particular: segmented filamentous bacteria (SFB), which make their home inside the small intestine of humans and other animals. Though these bacteria do not cause tissue damage or overt inflammation, they do stimulate an immune response in the host, resulting in the induction of immune cells that specifically recognize SFB and help control the population of these bacteria. This specificity relies on access of host cells to SFB antigens: foreign proteins or other distinctive components on bacterial targets that make them recognizable to immune cells. In the case of harmful bacteria, antigens arise in abundance as pathogens attack host cells and vice versa, but SFB don’t possess the machinery for invasion of host cells and show no hallmarks of destructive mechanisms. So how does the immune system come to recognize these peaceful residents? The answer, the scientists found, lies in a previously unknown communication pathway between commensal bacteria and host intestinal epithelial cells (IECs).

Using electron tomography, a technique that allows three-dimensional reconstruction of cellular and subcellular structures at high resolution, the authors found that IEC plasma membranes were forming small cavities exclusively where they interfaced with bacteria.  These cavities eventually bud off into bubble-like vesicles inside the host cell. This series of events at the host-bacteria interface was characteristic of normal cellular processes for bringing external substances into cells, known as endocytosis. Upon discovering that the observed vesicles contained a bacterial cell wall protein and common SFB antigen, the researchers confirmed that this pathway – which they termed “microbial adhesion-triggered endocytosis,” or “MATE” – served as a means by which SFB make their presence known to their host. Thus, host IECs can sample their commensal bacterial population without consuming and destroying whole microbial cells.  This peaceful transfer is likely advantageous in allowing the host to mount a mild immune response for SFB population control without triggering dramatic inflammation, though the mechanistic links between MATE and downstream immune effects remain unclear as of yet.

The authors, asking whether this unknown and surprisingly harmonious communication mechanism might be common among “healthy” microbes, next looked for signs of MATE among other classes of commensal intestinal bacteria, including those that are known to activate host immune responses similar to SFB responses. Though MATE communication was absent in all of the other species observed, the researchers noted that none of these microbes associated directly with IECs, as they observed in the case of SFB. Indeed, apart from SFB, the only microbes known to interact closely with IECs are bacterial pathogens, which themselves showed no signs of MATE signaling. These findings might indicate that MATE is a unique communication method specifically between host IECs and SFB (or other, as-yet-unidentified bacterial species), but they also suggest that strategies for crosstalk between microbes and hosts may be as diverse as the microbes themselves. 

Like MATE, many new pathways of host-commensal interaction might be awaiting discovery. Such pathways could someday open doors for alternative vaccine or drug-delivery strategies, reducing the necessity for much-dreaded needle shots. They may even facilitate therapies for regulating microbial populations as a revolutionary treatment for metabolic diseases like irritable bowel syndrome or obesity. If so, perhaps “germs” might get a little credit as heroes in the story of human health. Some good press at last.

 

Mark Ladinsky is an Electron Microscopy Scientist at the California Institute of Technology.  Dr. Leandro Araujo is a Postdoctoral Research Scientist in the Department of Microbiology & Immunology at Columbia University.

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