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Genomics of an Extreme Psychrophile, Psychromonas

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Genomics of an Extreme Psychrophile, Psychromonas BMC Genomics BioMed Central Research article Open Access Genomics of an extreme psychrophile, Psychromonas ingrahamii Monica Riley*1, James T Staley2, Antoine Danchin3, Ting Zhang Wang3, Thomas S Brettin4, Loren J Hauser5, Miriam L Land5 and Linda S Thompson4 Address: 1Bay Paul Center, Marine Biological Laboratory, Woods Hole, MA 02543, USA, 2University of Washington, Seattle, WA 98195-7242, USA, 3Genetics of Bacterial Genomes, CNRS URA2171, Institut Pasteur, 28 rue du Dr Roux, 75015 Paris, France, 4DOE Joint Genome Institute, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA and 5Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA Email: Monica Riley* - [email protected]; James T Staley - [email protected]; Antoine Danchin - [email protected]; Ting Zhang Wang - [email protected]; Thomas S Brettin - [email protected]; Loren J Hauser - [email protected]; Miriam L Land - [email protected]; Linda S Thompson - [email protected] * Corresponding author Published: 6 May 2008 Received: 3 September 2007 Accepted: 6 May 2008 BMC Genomics 2008, 9:210 doi:10.1186/1471-2164-9-210 This article is available from: http://www.biomedcentral.com/1471-2164/9/210 © 2008 Riley et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: The genome sequence of the sea-ice bacterium Psychromonas ingrahamii 37, which grows exponentially at -12C, may reveal features that help to explain how this extreme psychrophile is able to grow at such low temperatures. Determination of the whole genome sequence allows comparison with genes of other psychrophiles and mesophiles. Results: Correspondence analysis of the composition of all P. ingrahamii proteins showed that (1) there are 6 classes of proteins, at least one more than other bacteria, (2) integral inner membrane proteins are not sharply separated from bulk proteins suggesting that, overall, they may have a lower hydrophobic character, and (3) there is strong opposition between asparagine and the oxygen-sensitive amino acids methionine, arginine, cysteine and histidine and (4) one of the previously unseen clusters of proteins has a high proportion of "orphan" hypothetical proteins, raising the possibility these are cold-specific proteins. Based on annotation of proteins by sequence similarity, (1) P. ingrahamii has a large number (61) of regulators of cyclic GDP, suggesting that this bacterium produces an extracellular polysaccharide that may help sequester water or lower the freezing point in the vicinity of the cell. (2) P. ingrahamii has genes for production of the osmolyte, betaine choline, which may balance the osmotic pressure as sea ice freezes. (3) P. ingrahamii has a large number (11) of three-subunit TRAP systems that may play an important role in the transport of nutrients into the cell at low temperatures. (4) Chaperones and stress proteins may play a critical role in transforming nascent polypeptides into 3-dimensional configurations that permit low temperature growth. (5) Metabolic properties of P. ingrahamii were deduced. Finally, a few small sets of proteins of unknown function which may play a role in psychrophily have been singled out as worthy of future study. Conclusion: The results of this genomic analysis provide a springboard for further investigations into mechanisms of psychrophily. Focus on the role of asparagine excess in proteins, targeted phenotypic characterizations and gene expression investigations are needed to ascertain if and how the organism regulates various proteins in response to growth at lower temperatures. Page 1 of 19 (page number not for citation purposes) BMC Genomics 2008, 9:210 http://www.biomedcentral.com/1471-2164/9/210 Background Alamos, NM 87545. It was annotated at Oak Ridge Well over half of the earth's surface is cold: deep oceans, National Laboratory, Oak Ridge, TN 37831 and deposited mountains, polar regions. Likewise, Earth's solar system as GenBank file CP000510.1. Altogether 3708 genes were contains many planets and planetary bodies that are also identified, 3545 of which were proteins of 83 residues or cold. The cold environments on Earth are teeming with longer. A second round of annotation is described below. life [1] offering hope that other cold environments in our solar system such as Mars and Jupiter's moon, Europa, Properties of proteins may harbor life [2]. For this reason it is surprising that so Size little is know about the lifestyle, particularly of microbial One can ask whether the sizes of proteins of a psy- psychrophiles at low temperatures. chrophile differ from those of a mesophile. The sequences of all proteins of P. ingrahamii were compared to Psychrophiles have been studied primarily to understand sequences of all proteins of three other bacteria: biological mechanisms of adaptation to extreme condi- Shewanella oneidensis MR-1, Vibrio cholerae and Escherichia tions. Microbial physiologists have long been interested coli K-12 MG1655. 916 protein sequences were conserved in psychrophiles as they employ mechanisms allowing among all four bacteria. The great majority of the con- them to maintain life processes at temperatures where served proteins were enzymes. The distribution of lengths rates of reactions and molecular properties present chal- of the 916 orthologous proteins were compared, revealing lenges. In reaching for an understanding of how life proc- that the distribution was about the same for the compara- esses work at extremes of temperature, most of the focus ble proteins in all four bacteria (Figure 1). Ideas that pro- to date has been on the properties of enzymes of extrem- teins that are required to function at low temperatures ophiles (reviewed by [3,4]). No single consistent answer would be either shorter or longer than those of mes- has emerged to account for adaptation to temperature ophiles are not borne out. extremes. To date, no single type of modification is uni- formly found in the enzymes of psychrophiles; instead Amino acid composition numerous small and subtle differences appear to account Amino acid composition of an organism's proteins is for their increased flexibility thereby enabling them to affected by nucleotide composition of the DNA. The GC function at low temperatures. content of P. ingrahamii DNA was determined experimen- tally to be 40% [8], verified by the composition of the Recently whole genome sequences have been determined total genome nucleotide sequence (40.1%). Overall for psychrophiles Colwellia psychrerythraea 34 H [5], Idi- amino acid content of the encoded proteins compared to omarina loihiensis L2TR [6], and Pseudoalteromonas halo- those of V. cholerae, S. oneidensis and E. coli is shown in planktis TAC125 [7]. We now add the genomic sequence Table 1 (P. Sharp, personal comunication). The amino of the extreme species, Psychromonas ingrahamii 37 which acids isoleucine, asparagine, lysine, phenylalanine and grows at even colder temperatures. Availability of com- tyrosine all are present in higher percentage in P. ingra- plete genome sequences provides the opportunity to hamii. search all of the proteins of the organisms for similarities and differences that might have bearing on the ability of However, like the genome, the codons for these residues the organism to grow at low temperatures. are GC rich, a factor that must be taken into account. Codon usage for P. ingrahamii is shown in Table 2 for all The extreme psychrophile, Psychromonas ingrahamii was CDSs (coding DNA sequences) and for the highly isolated from sea ice from the Arctic. It grows exponen- expressed genes tuf, tsf, fusA+RP genes (P. Sharp, personal tially with a doubling time of 240 hours at -12°C and may communication). Thus when amino acid content is exam- well grow at even lower temperatures [8]. These tempera- ined as a function of GC3s, correcting for GC content at tures do not necessarily solidify salt water or cytoplasm synonymously variable third positions, the overall amino into ice. Liquid water has been shown to exist at grain con- acid composition of P. ingrahamii was found not to be tacts as low as -20C [2]. remarkable. At the value of 34.2 determined for P. ingra- hamii, amino acid contents fall on the curves generated for Results and Discussion GC3s dependency in 80 other organisms (P. Sharp, per- The P. ingrahamii genome sonal communication) (data not shown) [9]. The single, circular chromosome of 4.56 Mb constituting the genome of P. ingrahamii 37 was sequenced as a set of Correspondence analysis (CA) contigs by the DOE Joint Genome Institute Production Lobry and Chessel [10] examined datasets of amino acid Genomics Facility, 2800 Mitchell Drive, Walnut Creek, CA content and codon usage in thermophilic and mesophic 94598, and finished at DOE Joint Genome Institute, Bio- bacteria and found evidence that the amino acid compo- science Division, Los Alamos National Laboratory, Los sition of thermophilic proteins was under the control of a Page 2 of 19 (page number not for citation purposes) BMC Genomics 2008, 9:210 http://www.biomedcentral.com/1471-2164/9/210 100 80 60 E. coli S. oneidensis Number 40 V. cholerae Number P. ingrahamii 20 0 0 200 400 600 800 1000 1200 1400 1600 -20 Length (residues) Length (residues) DistributionFigure 1 of lengths of 916 orthologous proteins in four bacteria Distribution of lengths of 916 orthologous proteins in four bacteria. Distribution of lengths of proteins as numbers of amino acid residues, ranging from 83 to 1501, in increments of 30. Black diamond = E. coli, red square = S. oneidensis, yellow tri- angle = V. cholerae, blue cross = P. ingrahamii. pressure at the nucleic acid level, not a selection at the pro- We analyzed the amino acid compositions of all individ- tein level. An extended study [11] produced similar results ual proteins of P.
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