Identification of genes involved in the acetamidino group modification of the flagellin N-Linked glycan of Methanococcus maripaludis

N-linked glycosylation of protein is a posttranslational modification found in all three domains of life. The flagellin proteins of the archaeon Methanococcus maripaludis are known to be modified with an N-linked tetrasaccharide consisting of N-acetylgalactosamine (GalNAc), a diacetylated glucuronic acid (GlcNAc3NAc), an acetylated and acetamidino-modified mannuronic acid with a substituted threonine group (ManNAc3NAmA6Thr), and a novel terminal sugar residue [(5S)-2-acetamido-2,4-dideoxy-5-O-methyl-\u3b1-L-erythro-hexos-5-ulo-1,5-pyranose]. To identify genes involved in biosynthesis of the component sugars of this glycan, three genes, mmp1081, mmp1082, and mmp1083, were targeted for in-frame deletion, based on their annotation and proximity to glycosyltransferase genes known to be involved in assembly of the glycan. Mutants carrying a deletion in any of these three genes remained flagellated and motile. A strain with a deletion of mmp1081 had lower-molecular-mass flagellins in Western blots. Mass spectrometry of purified flagella revealed a truncated glycan with the terminal sugar absent and the threonine residue and the acetamidino group missing from the third sugar. No glycan modification was seen in either the \u394mmp1082 or \u394mmp1083 mutant grown in complex Balch III medium. However, a glycan identical to the \u394mmp1081 glycan was observed when the \u394mmp1082 or \u394mmp1083 mutant was grown under ammonia-limited conditions. We hypothesize that MMP1082 generates ammonia and tunnels it through MMP1083 to MMP1081, which acts as the amidotransferase, modifying the third sugar residue of the M. maripaludis glycan with the acetamidino group. \ua9 2012, American Society for Microbiology.Peer reviewed: YesNRC publication: Ye

Rest-Frame Mid-Infrared Detection of an Extremely Luminous Lyman Break Galaxy with the Spitzer Infrared Spectrograph (IRS)

We present the first rest-frame of approximately 4 microns detection of a Lyman break galaxy. The data were obtained using the 16 microns imaging capability of the Spitzer Infrared Spectrograph. The target object, J134026.44+634433.2, is an extremely luminous Lyman break galaxy at z=2.79, first identified in Sloan Digital Sky Survey (SDSS) spectra (as reported by Bentz et al.). The source is strongly detected with a flux of 0.94 +/- 0.02 mJy. Combining Spitzer and SDSS photometry with supporting ground-based J- and K-band data, we show that the spectral energy distribution is consistent with an actively star-forming galaxy. We also detect other objects in the Spitzer field of view, including a very red mid-infrared source. We find no evidence of a strong lens among the mid-infrared sources