Saturday, January 23, 2010

Mobilization and prevalence of a fusobacterial plasmid

Mobilization and prevalence of a fusobacterial plasmid.
Brianna M. Claypool, Sean C. Yoder, Diane M. Citron, Sydney M. Finegold, Ellie J.C. Goldstein and Susan Kinder Haake

Plasmid 2010 63:11-19

Fusobacterium nucleatum is a Gram-negative anaerobic rod found in dental plaque biofilms, and is an opportunistic pathogen implicated in periodontitis as well as a wide range of systemic abscesses and infections.
Studies indicate considerable phenotypic variability between F. nucleatum strains, and biochemical analyses have suggested that F. nucleatum represents a “species complex”. Comparative genomic analyses revealed that 25% of the genes encoded by F. nucleatum ATCC 10953 ssp. polymorphum are not found in either of the sequenced genomes of F. nucleatum ssp. nucleatum and vincentii). In addition, 21% of these unique ORFs mapped to clusters of five or more genes, suggesting that they may have been introduced into the genome by horizontal gene transfer. Several plasmids were found in Fusobacterium but they are too small to be self transmissible.
In this paper authors defined the minimal “mobilon” of plasmid pFN1 that included the relaxase, and DNA directly upstream containing ORF4 and ORF7 with the oriT region located upstream of ORF4. The pFN1 mobilon is related by sequence to the mobilons of the staphylococcal plasmids pC221 and pC223. Authors screened 94 clinical and 4 laboratory isolates of F. nucleatum for plasmids and plasmid encoded relaxase gene and they found that 11.5% of isolates.
As the main discovery authors demonstrated for the first time that the fusobacterial plasmid pFN1 encodes genes enabling efficient mobilization between strains of E. coli by the broad-host range plasmid RP4. I found this experiment really interesting. It is true that IncP plasmid RP4 can mobilize plasmid pFN1 between E. coli strains but if we look closer we can find that plasmid pFN1 cannot replicate in E. coli as oriP15A was used for stable replication. On the other hand there is no evidence that pRP4 can transfer to and/or replicate in F. nucleatum. This can raise a question, how is it possible for pRP4 to mobilize the pFN1 plasmid in vivo. Unfortunately, authors did not discuss that point. On the other hand they realize that the main aim to really prove the role of Fusobacterium plasmids in horizontal gene transfer is to show that plasmid transfer occurs in vivo and find what can support the missing conjugation functions.
Our dual reporter system for detection plasmid transfer in situ in anaerobic conditions can help in this research.

Jarek Krol

Tuesday, January 12, 2010

Into the Woods

The fundamental units, processes and patterns of evolution, and the Tree of Life conundrum

Eugene V Koonin and Yuri I.Wolf

Biology Direct (2009), 4:33

Human communication and thought is largely shaped by metaphors. Metaphors allow man to think about concepts and facts in new ways and to make connections that otherwise might have been left undiscovered. However, if one becomes too committed to a particular metaphor there is the danger of the opposite-of stifling creativity and having a distorted view of individual facts or even the world at large.

"The affinities of all the beings of the same class have sometimes been represented by a great tree. I believe this simile largely speaks the truth. The green and budding twigs may represent existing species; and those produced during each former year may represent the long succession of extinct species. .... The limbs divided into great branches, and these into lesser and lesser branches, were themselves once, when the tree was small, budding twigs; and this connexion of the former and present buds by ramifying branches may well represent the classification of all extinct and living species in groups subordinate to groups."

The authors of this article begin with this quote from Charles Darwin(1) in order to assert that the very roots of evolutionary theory are grounded in the metaphor of a “Tree of Life.” In this metaphor all living organisms are the budding tips of living branches in the Tree of Life (TOL), whose trunk was the original organism from which all life derived. While this metaphor has served scientists well for centuries, our increased understanding of horizontal gene transfer has led to a “crisis of the TOL.”

The authors argue that individual viruses, plasmids, transposons, individual genes, and the like, rather than organisms or species, should be considered as the true fundamental units of evolution (FUEs). The evolutionary history of each individual FUE is still accurately represented by a tree in that an ancestral gene can be altered such that a new version “branches off,” with this process continuing until a full tree is formed. An organism, therefore, is made up of many such FUE trees, and so is more properly a “forest of life.”

The more we learn about the importance of horizontal gene transfer among organisms, species, or even kingdoms, the more clear it becomes that the metaphor of a Tree of Life ignores much of the complexity of true evolutionary histories, especially among prokaryotes. We therefore might benefit from reevaluating our metaphor and perhaps, as the authors suggest, acknowledge the true complexity of the Forest of Life.

References\Further Reading:

1. Darwin C: On the Origin of Species. 1st edition. London: Murray; 1859.

2. Hilario E, Gogarten JP: Horizontal transfer of ATPase genes--the tree of life becomes a net of life. Biosystems31(2-3):111-119. 1993,

3. Doolittle WF: Uprooting the tree of life. Sci Am 2000, 282(2):90-95

4. Nelson KE, Clayton RA, Gill SR, Gwinn ML, Dodson RJ, Haft DH, Hickey EK, Peterson JD, Nelson WC, Ketchum KA, et al.: Evidence for lateral gene transfer between Archaea and bacteria from genome sequence of Thermotoga maritima. Nature 1999, 399(6734):323-329.

Julie M. Hughes

University of Idaho