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SMALL THINGS CONSIDERED A BLOG FOR SHARING APPRECIATION OF THE WIDTH AND DEPTH OF MICROBES AND MICROBIAL ACTIVITIES ON THIS PLANET. Small Things Considered * About * Contact * Subscribe « All the World Is Not E. coli | Main | Prokaryotic Organelles. Yes, There Are Such Things. » PICTURES CONSIDERED #52 : STREPTOMYCES STACKS Z-RINGS... by Christoph Figure 1. Time-lapse fluorescence and DIC microscopy movie showing the localization of FtsZ-YPet in vegetative and sporulating hyphae of wild-type S. venezuelae (SS12). Note the timer in the upper right corner. Source Streptomyces stacks Z-rings... into ladders inside its aerial/sporogenic hyphae. If this sentence strikes you as a barrage of jargon or just gibberish, don't despair, help is coming. First, enjoy the "light show" in the 20‑second video clip on the right side. It runs in loop, so you can follow more than just one growing hypha, and see what happens in them over time (Figure 1). Click on the picture to see an enlarged version in a separate window. Now taking apart the first sentence: on Petri dishes, Actinobacteria of the genus Streptomyces grow as long rod-like hyphae (filaments) with infrequent septa and occasional branches that reach into the substrate and protrude into the air (see Figure 2). The aerial or sporogenic hyphae differentiate into chains of spores, which break-off when mature (maybe they are taken on a ride by passing Bacillus subtilis, as described here in STC, who knows.) The differentiation process involves the action of FtsZ, the cytoskeletal protein that forms constricting rings attached to the cytosolic side of the membrane and drives cell septation and division. The video clip is part of a recent study by Susan Schlimpert's lab at the John Innes Centre, Norwich, UK. "Starring" in the video clip is Streptomyces venezuelae, which unlike most other Streptomyces species sporulates submersed in liquid media, and Susan provided a few more details in an email: "The movie covers a period of ~11 hours (1 frame/10 min). We usually cut out the first 6–8 h of the lifecycle and only start imaging around the time when we expect sporulation to happen. We grow Streptomyces venezuelae (or Sven called in the lab) in a commercial microfluidics system which allows us to flush-in "spent medium" (medium from a sporulating culture). This stimulates sporulation. One spore-to-spore lifecycle takes about 24h." It is far from trivial to "tag" a protein capable of oligomerization with a yellow fluorescent protein (YFP) without affecting its intracellular localization and functionality, but it worked in this case without the cells needing another "untagged" gene copy. Susan wrote in an earlier methods paper: "In sporulating hyphae, the localization pattern of FtsZ-YPet changes dramatically; first helical FtsZ-YPet filaments tumble along the hypha and then, in a sudden, almost synchronous event, these helices coalesce into a ladder of regularly spaced FtsZ-YPet rings. Under the experimental conditions described here, these evenly distributed FtsZ-YPet ladders persist for approximately 2 hr. Finally, sporulation septa become discernible in the differential interference contrast (DIC) images... and eventually new spores are released." Figure 2. Streptomyces life-cycle on solid media. The cellular development of a spore begins with the formation of one or two germ tubes, which grow by tip extension to form a network of branching hyphae. Polar growth and branching of the vegetative hyphae is directed by DivIVA (red). The formation of vegetative cross-walls requires FtsZ (green). In response to nutrient limitations and other signals, aerial hyphae are erected. Arrest of aerial growth is tightly coordinated with the assembly of a ladder of FtsZ-rings, which give rise to the sporulation septa that compartmentalize the sporogenic hyphae into box-like prespore compartments. These compartments assemble a thick spore wall and are eventually released as mature pigmented spores. Source It was actually not the aim of Ramos-León et al. to obtain the longest and most beautiful Z-ring ladders but rather to use them as assay system to get clues about the function of a protein, SepH, that is (so far) only found in Actinobacteria. Indeed, they found that a S. venezuelae ΔsepH mutant shows aberrant Z-ring placement in the sporogenic hyphae, and the resulting spores are highly irregular in size. SepH binds FtsZ and stimulates the assembly of FtsZ protofilaments in vitro. It appears that SepH promotes the bundling of FtsZ protofilaments in the correct inter-filament distances to achieve Z-ring condensation in vivo, indicating a Z-ring stabilizing function as is known for the FtsZ-binding proteins (ZBPs) EzrA, SepF and ZapA in B. subtilis. This is not the first time we feature a short video clip in our "Pictures Considered" series (#15 was the first), and we'll be sure to do so more often in the future. Not because it is now technically possible − and entertaining − but because 'motion pictures' capture the inherent dynamics of biological processes in a way that is superior to still photographs for their understanding, "telling" as you may call it. This is an ever‑so‑slight contradiction to Elio's statement "One picture is all it took," when he celebrated "The Birth of the FtsZ Ring" in Pictures Considered #5. However, in 1991 when Erfei Bi & Joe Lutkenkaus presented the first images of FtsZ rings, observing and filming proteins in living bacterial cells was still in its infancy. To my knowledge, the time-lapse series of oscillating GFP-tagged MinD protein in E. coli, shown by Piet de Boer during a workshop I attended in 1997, were "a first." Posted on May 06, 2021 at 01:30 AM in Pictures Considered, Teachers Corner | Permalink COMMENTS VERIFY YOUR COMMENT PREVIEWING YOUR COMMENT Posted by: | This is only a preview. Your comment has not yet been posted. Your comment could not be posted. Error type: Your comment has been saved. Comments are moderated and will not appear until approved by the author. Post another comment The letters and numbers you entered did not match the image. Please try again. As a final step before posting your comment, enter the letters and numbers you see in the image below. This prevents automated programs from posting comments. Having trouble reading this image? View an alternate. POST A COMMENT Comments are moderated, and will not appear until the author has approved them. Comment below or sign in with Typepad Facebook Twitterand more... You are currently signed in as (nobody). Sign Out Your Information (Name is required. Email address will not be displayed with the comment.) Name is required to post a comment Please enter a valid email address Invalid URL Please enable JavaScript if you would like to comment on this blog. Small Things Considered Links About Contact Subscribe FAQ Archives Elio's Memoirs In Memoriam Our Books etc. Social Media Links Instagram 𝕏 (Twitter) Mastodon Bluesky ASM Links American Society for Microbiology About ASM ASM Publications Join ASM This Week in Microbiology (TWiM) ©2023 Small Things Considered Privacy Policy, Terms of Use, and State Disclosures