Posts Tagged ‘Cell Biology’

Implanted Genome

Posted by amanda on Friday, May 21st, 2010

Mycoplasma

Mycoplasma

Is a rose still a rose if it doesn’t smell as sweet? Is a Mycoplasma capricolum still a Mycoplasma capricolum if it expresses a different genome? The answer to the first question is definitely “Yes” (especially if you could smell the roses I received on Mother’s Day). A paper published yesterday in Science confirms that the answer to the second question is a big fat “No”. Researchers at Maryland’s own J. Craig Venter Institute (JCVI) in Rockville in collaboration with their sister institute in San Diego are the first to successfully implant a new genome into a bacterial cell to create a new self-replicating bacteria that never existed before. In the most simplistic description, they created a unique new life.

Using a novel technique in which large DNA sequences were strung together in yeast, a 1.08 Mbp (Mega base pairs, where a base pair is one unit or nucleotide of double-stranded DNA) encoding all the necessary genes to make Mycoplasma mycoides JCVI-syn1.0 was synthesized. After synthesis, the circular chromosome was implanted into the nucleus of a similar species, Mycoplasma capricolum, for gene expression. This was not easy. After the first attempt failed, the researchers found a single mutation limiting proper gene expression. In their paper, the scientists recall several obstacles that had to be overcome: They needed to develop a method to extract large chromosomes from yeast, learn how to transplant these genomes into a recipient cell, and to choose a fast-growing recipient with the required gene expression machinery as a donor.

By synthesizing the DNA themselves, the researchers were able to include “markers” within the sequence. For example, if the genes were successfully expressed, the bacteria would be blue in color. Furthermore, they were able to leave a lasting mark on the DNA sequence by including strings of nucleotides (that when expressed into protein) spelled out an email address, the names of the scientists involved in the project, and a few famous quotations. [It isn't clear whether or not these proteins would actually be expressed. If only you could make an enzyme composed of a Shakespeare quote.]

This paper has raised some ethical and therefore political issues. President Obama has already asked the White House bioethics commission for a review of the issues to be provided to him in a report within 6 months, stating that the creation of a new genome raised “genuine concerns”. The application of this method for the synthesis of novel compounds using bacteria, such as biofuels, make this an important industrial topic as well. Can you patent an organism that you created? Would the laws that apply to genetically engineered crops where patents are in place apply to this as well?

In the last paragraph of the paper, the researchers write:

“We have been driving the ethical discussion concerning
synthetic life from the earliest stages of this work (25, 26). As
synthetic genomic applications expand, we anticipate that this
work will continue to raise philosophical issues that have
broad societal and ethical implications. We encourage the
continued discourse.”

Let the discourse begin.

Heat-sensing Vipers

Posted by amanda on Thursday, March 18th, 2010

pit viper

Timber rattlesnake; Taken by Tad Arensmeier from St. Louis, MO, USA

Hikers in Maryland are repeatedly warned at trailheads about the presence of venomous snakes. According to the Maryland DNR, there are over 27 species and subspecies of snakes in Maryland, including two venomous pit vipers, the copperhead and timber rattlesnake. Pit vipers are named for their large specialized pit organs located between the nostril and eye on each side of their face. These organs allow pit vipers to sense infrared radiation, detecting warm-blooded prey at temperatures above ~30°C and distances of up to one meter.

The pit organ is composed of a thin membrane suspended in a hollow chamber. This membrane, serving as an infrared antenna, is packed with mitochondria (cellular power plants) and linked to many nerve fibers. These fibers send signals from the pit organs to the brain. The sensitivity of detection is dependent on the anatomical and molecular characteristics of the pit organ.

This week in a Nature advance online publication, scientists reveal the identity of the molecular sensor used by pit vipers and describe how these sensors convey information to the snake brain. Rather than “seeing” in infrared, the pit organ actually heats up due to the ion channel TRPA1. Researchers found TRPA1 in nerve endings leading to the pit organ at levels 400x that in other nerve endings. Humans and other mammals also have TRPA1 (63% similar to pit viper) which is activated by the pungent agent from wasabi and mustard plants. Unlike our TRPA1, the pit viper receptor is not only activated by the mustard agent but also by heat above 28°C. The heat activation of these channels provides an infrared map to the snake brain.

Interestingly, the researchers found no activation of TRPA1 channels in cooler temperatures, although it is known that snakes can respond to temperatures well below 28°C, perhaps to locate cooler areas in which to rest. Is there another receptor that can detect cooler climes? Or maybe the anatomy of the pit organ tunes TRPA1 in a way we haven’t yet explored.

According to the Maryland DNR, only 2-5 people get bitten by a venomous snake in the state each year. If you happen to be one of the unlucky ones, just remember:
- Don’t apply ice.
- Do keep the bite immobilized and below your heart.
- Don’t apply a tourniquet.
- Do wash it with soap and water.
- Don’t make an incision.
- Do get help immediately.

The Internal Compass

Posted by amanda on Thursday, March 11th, 2010

Billions of white blood cells are streaming through your bloodstream. They’re on patrol, defending you from foreign invaders and protecting you from infection. Some respond to parasites, others to viruses and allergens. When certain white bloods cells called neutrophils detect an infection or area of injury, they will move in, engulfing and digesting any foreign bacteria or fungi in their path. But how does a neutrophil know where to go? How does it sense its target? What is its internal compass?

Neutrophils, and many other cells, use chemotaxis, the detection and directed movement of cells in response to a chemical gradient. In an advance publication of the Annual Review of Biophysics, Kristen Swaney and members of the Devreotes Lab in the Department of Cell Biology at Johns Hopkins University review the process of chemotaxis in eukaryotic cells (like our neutrophils). Much of what we know about chemotaxis in these cells is from the study of Dictyostelium, a soil-living amoeba commonly (and lovingly?) known as slime mold.

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Neutrophil chasing a bacterium

The process of chemotaxis does not actually begin with signal detection but with random cell movement. Cells like neutrophils and Dictyostelium extend pseudopodia (literally “fake feet”) made up of filaments consisting of many molecules of actin protein. The actin in these filaments interacts with myosin protein at the cell’s end, causing contraction (just like in your muscle cells), propelling the cell forward. After extension, the cell reassembles the actin filaments, so it can begin again. With this ambling gait, the cell moves in a random direction. In the absence of an attractant, cells extend pseudopodia from each side uniformly. But once the cell senses an attractant, pseudopodia extend toward the signal. Most notable is the wave-like motion through the cell when an attractant is present. This wave is caused from the recruitment of actin-binding proteins sequentially from the inside of the cell to points on the surface and may underlie the generation of pseudopodia.

Cells detect chemical attractants with receptors on their surfaces. Once receptors bind an attractant molecule, a signal cascade is initiated and a network of signaling pathways is activated leading to the formation of actin into filaments and movement. Eukaryotic cells are capable of sensing both the amount of attractant and the location of the attractant in space, detecting how many receptors have bound attractant and where those receptors are located on their surface. Interestingly, if an attractant is present in low amounts, the cell will “adapt” to the presence of attractant and actually be more sensitive to an attractant gradient by ignoring the background.

Genetic analysis of Dictyostelium has revealed a complex and wide range of interacting players in the process of chemotaxis. In her review, Swaney emphasizes that we still have much to learn about the components involved in wave propagation, the mechanism of attractant adaptation, the localization of components, and the complexity of multiple signaling networks. There are many pieces to an internal compass.