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28.02.2012 In­trons found in bac­terial 16S rRNA

Max Planck sci­ent­ists from Bre­men identify long pieces of non-cod­ing DNA in the most im­port­ant phylo­gen­etic marker gene in bac­teria
 
The char­ac­ter­istic that mi­cro­bi­o­lo­gists use to de­term­ine the phylo­gen­etic af­fil­i­ation in bac­teria needs to meet cer­tain de­mands. It has to be con­ser­vat­ive in its func­tion and present in all or­gan­isms. A cer­tain gene meets this re­quire­ment to a high de­gree: the gene for the smal­ler one of the two sub­units that to­gether form the ri­bo­some, also known as 16S rRNA gene. For that reason it is fre­quently used as phylo­gen­etic marker in ana­lyses that re­search­ers per­form to de­term­ine the de­gree of re­lated­ness in mi­crobes.

Ver­ena Sal­man from the Max Planck In­sti­tute for Mar­ine Mi­cro­bi­o­logy in Bre­men has, to­gether with her col­leagues, taken a closer look at the 16S rRNA gene of a cer­tain bac­terial group, the large sul­fur bac­teria (Fig­ure 1). They found out that the 16S rRNA gene of some mem­bers of this group is in­ter­rup­ted by long DNA in­serts at up to four sites (Fig­ure 2). However, these so called in­trons are not DNA in­serts that en­code for 16S rRNA: They either con­sist of non-cod­ing DNA or they carry genes for cer­tain en­zymes by which the in­tron can spread into other parts of the gen­ome.
Figure 1: The cells of the large sulfur bacteria Thiomargarita often occur in chains. In their 16S rRNA gene the scientists found four introns. At a diameter of 200 μm the cells are extremely large for bacterial cells. Image: V. Salman
“Each in­tron has a length of sev­eral hun­dred base pairs. Hence, they in­crease the length of the 16S rRNA gene con­sid­er­ably. We found 16S rRNA genes of up to 3500 base pairs length”, says Ver­ena Sal­man. The com­mon 16S rRNA gene is only 1500 base pairs long. The sci­ent­ists suc­ceeded for the first time to prove the ex­ist­ence of these in­trons in the marker gene of bac­terial 16S rRNA. This is sur­pris­ing, since this gene is the most se­quenced gene today.

The dis­cov­ery of the in­trons in the 16S rRNA gene has sub­stan­tial im­plic­a­tions for the mi­cro­bi­o­lo­gists. “So far, we could only se­quence short pieces of the 16S rRNA genes of the large sul­fur bac­teria from a mixed bac­terial sample. Now we know why: Their 16S rRNA gene is longer than that of other bac­teria,” ex­plains Rudolf Amann, dir­ector of the Max Planck In­sti­tute. “Mi­cro­bial cells that carry the long ver­sion of the gene can­not be iden­ti­fied by the clas­sical meth­ods, be­cause the isol­a­tion of shorter se­quences is strongly pre­ferred through these meth­ods.” For that reason the sci­ent­ists could so far not de­tect the gene in its full length (Fig­ure 3).
Figure 2: In the schematic view of the 16S rRNA gene the positions of the introns are marked in light grey. The introns are removed before the genetic information of the ribosome is read. Image: V. Salman/R. Dunker
Since the 16S rRNA gene plays an im­port­ant role in the phylo­gen­etic clas­si­fic­a­tion of bac­teria and in biod­iversity stud­ies, the dis­cov­ery of the sci­ent­ists from Bre­men has far-reach­ing con­sequences. Pre­sum­ably, the in­trons also oc­cur in the 16S rRNA genes of other bac­terial groups. “Cur­rently we try to find out which other mi­croor­gan­isms also carry the elong­ated 16S rRNA gene”, as Ver­ena Sal­man says. “With this in­form­a­tion we will de­velop a strategy that also con­siders the long 16S rRNA se­quences, for in­stance in biod­iversity stud­ies”.
The bac­teria do not seem to be af­fected by the in­trons in the 16S rRNA genes. Des­pite the long in­trons the cells of the large sul­fur bac­teria de­velop func­tional ri­bosomes. The in­trons re­move them­selves dur­ing the ri­bo­some form­a­tion pro­cess (Fig­ure 2).
Figure 3: 16S rRNA-genes from bacteria in a mixed sample can only be detected by the standard methods if they belong to a certain length scheme. Longer 16S rRNA genes, e.g. those with inserts, remain undetected and hence are missing in biodiversity studies. The normal 16S rRNA gene is marked in black, the elongated version in red. Image: V. Salman/R. Dunker
For fur­ther in­form­a­tion please con­tact

Dr. Ver­ena Sal­man
Prof. Dr. Heide Schulz-Vogt
Prof. Dr. Rudolf Amann

Or the pub­lic re­la­tions of­fice
Dr. Rita Dunker
Dr. Man­fred Schlösser

Ori­ginal art­icle

Mul­tiple self-spli­cing in­trons in the 16S rRNA genes of gi­ant sul­fur bac­teria, 2012. V. Sal­man, R. Amann, D. A. Shub, and H. Schulz-Vogt. Pro­ceed­ings of the Na­tional Academy of Sci­ence, Early Edi­tion.

DOI: 10.1073/​pnas.1120192109

In­volved in­sti­tu­tions

Max Planck In­sti­tute for Mar­ine Mi­cro­bi­o­logy, Bre­men, Ger­many
Uni­versity of Al­bany, State Uni­versity of New York, Al­bany, USA
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