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Journal of Bacteriology, May 2009, p. 3068-3075, Vol. 191, No. 9
0021-9193/09/$08.00+0     doi:10.1128/JB.01838-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Manual Annotation, Transcriptional Analysis, and Protein Expression Studies Reveal Novel Genes in the agl Cluster Responsible for N Glycosylation in the Halophilic Archaeon Haloferax volcanii{triangledown} ,{dagger}

Sophie Yurist-Doutsch and Jerry Eichler*

Department of Life Sciences, Ben Gurion University, Beersheva 84105, Israel

Received 31 December 2008/ Accepted 14 February 2009

While Eukarya, Bacteria, and Archaea are all capable of protein N glycosylation, the archaeal version of this posttranslational modification is the least understood. To redress this imbalance, recent studies of the halophilic archaeon Haloferax volcanii have identified a gene cluster encoding the Agl proteins involved in the assembly and attachment of a pentasaccharide to select Asn residues of the surface layer glycoprotein in this species. However, because the automated tools used for rapid annotation of genome sequences, including that of H. volcanii, are not always accurate, a reannotation of the agl cluster was undertaken in order to discover genes not previously recognized. In the present report, reanalysis of the gene cluster that includes aglB, aglE, aglF, aglG, aglI, and aglJ, which are known components of the H. volcanii protein N-glycosylation machinery, was undertaken. Using computer-based tools or visual inspection, together with transcriptional analysis and protein expression approaches, genes encoding AglP, AglQ, and AglR are now described.

* Corresponding author. Mailing address: Dept. of Life Sciences, Ben Gurion University, P.O. Box 653, Beersheva 84105, Israel. Phone: (972) 8646 1343. Fax: (972) 8647 9175. E-mail: jeichler{at}bgu.ac.il

{triangledown} Published ahead of print on 27 February 2009.

{dagger} Supplemental material for this article is available at http://jb.asm.org/.

Journal of Bacteriology, May 2009, p. 3068-3075, Vol. 191, No. 9
0021-9193/09/$08.00+0     doi:10.1128/JB.01838-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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