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61.
Garry W. Buchko Jan Abendroth John I. Robinson Isabelle Q. Phan Peter J. Myler Thomas E. Edwards 《Protein science : a publication of the Protein Society》2020,29(3):670-685
A protein superfamily with a “Domain of Unknown Function,”, DUF3349 (PF11829), is present predominately in Mycobacterium and Rhodococcus bacterial species suggesting that these proteins may have a biological function unique to these bacteria. We previously reported the inaugural structure of a DUF3349 superfamily member, Mycobacterium tuberculosis Rv0543c. Here, we report the structures determined for three additional DUF3349 proteins: Mycobacterium smegmatis MSMEG_1063 and MSMEG_1066 and Mycobacterium abscessus MAB_3403c. Like Rv0543c, the NMR solution structure of MSMEG_1063 revealed a monomeric five α‐helix bundle with a similar overall topology. Conversely, the crystal structure of MSMEG_1066 revealed a five α‐helix protein with a strikingly different topology and a tetrameric quaternary structure that was confirmed by size exclusion chromatography. The NMR solution structure of a fourth member of the DUF3349 superfamily, MAB_3403c, with 18 residues missing at the N‐terminus, revealed a monomeric α‐helical protein with a folding topology similar to the three C‐terminal helices in the protomer of the MSMEG_1066 tetramer. These structures, together with a GREMLIN‐based bioinformatics analysis of the DUF3349 primary amino acid sequences, suggest two subfamilies within the DUF3349 family. The division of the DUF3349 into two distinct subfamilies would have been lost if structure solution had stopped with the first structure in the DUF3349 family, highlighting the insights generated by solving multiple structures within a protein superfamily. Future studies will determine if the structural diversity at the tertiary and quaternary levels in the DUF3349 protein superfamily have functional roles in Mycobacteria and Rhodococcus species with potential implications for structure‐based drug discovery. 相似文献
62.
There are gaps in existing understanding of fungal pellet growth dynamics. We used scanning electron microscopy (SEM) for morphological characterization of the biomass organization of Termitomyces pellets for seven species: T. microcarpus (TMI1), T. albuminosus (TAL1, TAL2), T. striatus (TSTR), T. aurantiacus (TAUR), T. heimii (THE1, THE2), T. globulus (TGLO) and T. clypeatus (TCL1, TCL2, TCL3, TCL4, TCL5). We assessed the utility of SEM for morphological and structural characterization of Termitomyces spp. in three dimensional (3D) pellet form to identify ideal pellet morphology for industrial use. Typological classification of Termitomyces species was based on furrows, isotropy, total motifs and fractal dimensions. The pellets formed were entangled and exhibited highly compacted mycelial mass with microheterogeneity and microporosity. The mean density of furrows of Termitomyces species was between 10,000 and 11,300 cm/cm2, percentage isotropy was 30?80 and total motifs varied from 300 to 2500. TGLO exhibited the highest furrow mean density, 11243 cm/cm2, which indicated a compact, cerebroid structure with complex ridges and furrows, whereas TAL2 exhibited the lowest furrow density. TMI1a exhibited a high percentage isotropic value, 74.6, TSTR exhibited the lowest, 30.9. Total motif number also was used as a typological classification parameter. Fractal values were 2.64?2.78 for various submerged conditions of Termitomyces species. TAL1 exhibited the highest fractal dimension and TAL2 the lowest, which indicates the complexity of branching patterns. Three-dimensional SEM image analysis can provide insight into pellet micromorphology and is a powerful tool for exploring topographical details of pellets. 相似文献
63.
Unusual pattern of bacterial ice nucleation gene evolution 总被引:5,自引:0,他引:5
Edwards AR; Van den Bussche RA; Wichman HA; Orser CS 《Molecular biology and evolution》1994,11(6):911-920
Bacterial ice nucleation activity (INA+ phenotype) can be traced to the
product of a single gene, ina. A remarkably sparse distribution of this
phenotype within three bacterial genera indicates that the ina gene may
have followed an unusual evolutionary path. Southern blot analyses, coupled
with assays for ice-nucleating ability, revealed that within four bacterial
species an ina gene is present in some strains but absent from others.
Results of hybridization experiments using DNA fragments that flank the ina
gene suggested that the genotypic dimorphism of ina may be anomalous. A
phylogenetic analysis of 16S ribosomal RNA gene sequences from a total of
14 ina+ and ina- bacterial strains indicated that the ina+ bacteria are not
monophyletic but instead phylogenetically interspersed among ina- bacteria.
The relationships of ina+ bacteria inferred from ina sequence did not
coincide with those inferred from the 16S data. These results suggest the
possibility of horizontal transfer in the evolution of bacterial ina genes.
相似文献
64.
65.
Association of Guide RNA Binding Protein gBP21 with Active RNA Editing Complexes in Trypanosoma brucei 总被引:2,自引:0,他引:2 下载免费PDF全文
Thomas E. Allen Stefan Heidmann RoseMary Reed Peter J. Myler H. Ulrich Gringer Kenneth D. Stuart 《Molecular and cellular biology》1998,18(10):6014-6022
RNA editing in Trypanosoma brucei mitochondria produces mature mRNAs by a series of enzyme-catalyzed reactions that specifically insert or delete uridylates in association with a macromolecular complex. Using a mitochondrial fraction enriched for in vitro RNA editing activity, we produced several monoclonal antibodies that are specific for a 21-kDa guide RNA (gRNA) binding protein initially identified by UV cross-linking. Immunofluorescence studies localize the protein to the mitochondrion, with a preference for the kinetoplast. The antibodies cause a supershift of previously identified gRNA-specific ribonucleoprotein complexes and immunoprecipitate in vitro RNA editing activities that insert and delete uridylates. The immunoprecipitated material also contains gRNA-specific endoribonuclease, terminal uridylyltransferase, and RNA ligase activities as well as gRNA and both edited and unedited mRNA. The immunoprecipitate contains numerous proteins, of which the 21-kDa protein, a 90-kDa protein, and novel 55- and 16-kDa proteins can be UV cross-linked to gRNA. These studies indicate that the 21-kDa protein associates with the ribonucleoprotein complex (or complexes) that catalyze RNA editing. 相似文献
66.
Thomas E. Edwards Isabelle Phan Jan Abendroth Shellie H. Dieterich Amir Masoudi Wenjin Guo Stephen N. Hewitt Angela Kelley David Leibly Mitch J. Brittnacher Bart L. Staker Samuel I. Miller Wesley C. Van Voorhis Peter J. Myler Lance J. Stewart 《PloS one》2010,5(9)
Background
Pathogenic bacteria adhere to the host cell surface using a family of outer membrane proteins called Trimeric Autotransporter Adhesins (TAAs). Although TAAs are highly divergent in sequence and domain structure, they are all conceptually comprised of a C-terminal membrane anchoring domain and an N-terminal passenger domain. Passenger domains consist of a secretion sequence, a head region that facilitates binding to the host cell surface, and a stalk region.Methodology/Principal Findings
Pathogenic species of Burkholderia contain an overabundance of TAAs, some of which have been shown to elicit an immune response in the host. To understand the structural basis for host cell adhesion, we solved a 1.35 Å resolution crystal structure of a BpaA TAA head domain from Burkholderia pseudomallei, the pathogen that causes melioidosis. The structure reveals a novel fold of an intricately intertwined trimer. The BpaA head is composed of structural elements that have been observed in other TAA head structures as well as several elements of previously unknown structure predicted from low sequence homology between TAAs. These elements are typically up to 40 amino acids long and are not domains, but rather modular structural elements that may be duplicated or omitted through evolution, creating molecular diversity among TAAs.Conclusions/Significance
The modular nature of BpaA, as demonstrated by its head domain crystal structure, and of TAAs in general provides insights into evolution of pathogen-host adhesion and may provide an avenue for diagnostics. 相似文献67.
Transgene-mediated and elicitor-induced perturbation of metabolic channeling at the entry point into the phenylpropanoid pathway 总被引:12,自引:0,他引:12 下载免费PDF全文
3H-l-Phenylalanine is incorporated into a range of phenylpropanoid compounds when fed to tobacco cell cultures. A significant proportion of (3)H-trans-cinnamic acid formed from (3)H-l-phenylalanine did not equilibrate with exogenous trans-cinnamic acid and therefore may be rapidly channeled through the cinnamate 4-hydroxylase (C4H) reaction to 4-coumaric acid. Such compartmentalization of trans-cinnamic acid was not observed after elicitation or in cell cultures constitutively expressing a bean phenylalanine ammonia-lyase (PAL) transgene. Channeling between PAL and C4H was confirmed in vitro in isolated microsomes from tobacco stems or cell suspension cultures. This channeling was strongly reduced in microsomes from stems or cell cultures of transgenic PAL-overexpressing plants or after elicitation of wild-type cell cultures. Protein gel blot analysis showed that tobacco PAL1 and bean PAL were localized in both soluble and microsomal fractions, whereas tobacco PAL2 was found only in the soluble fraction. We propose that metabolic channeling of trans-cinnamic acid requires the close association of specific forms of PAL with C4H on microsomal membranes. 相似文献
68.
69.
70.
Phylogenetic analyses of the rbcL sequences from haptophytes and heterokont algae suggest their chloroplasts are unrelated 总被引:2,自引:0,他引:2
Using the large subunit of RuBisCo (rbcL) sequences from cyanobacteria,
proteobacteria, and diverse groups of algae and green plants, we evaluated
the plastid relationship between haptophytes and heterokont algae. The rbcL
sequences were determined from three taxa of heterokont algae
(Bumilleriopsis filiformis, Pelagomonas calceolata, and Pseudopedinella
elastica) and added to 25 published sequences to obtain a data set
comprising 1,434 unambiguously aligned sites (approximately 98% of the
total rbcL gene). Higher levels of mutational saturation in third codon
positions were observed by plotting the pairwise substitutions with and
without corrections for multiple substitutions at the same site for first
and second codon positions only and for third positions only. In accordance
with this finding phylogeny reconstructions were completed by omitting
third codon positions, thus using 956 bp in weighted-parsimony and
maximum-likelihood analyses. The midpoint-rooted phylogenies showed two
major clusters, one containing cyanobacteria, glaucocystophytes, a
phototrophic euglenoid, chlorophytes, and embryophytes (the green lineage),
the other containing proteobacteria, haptophytes, red algae, a cryptophyte,
and heterokont algae (the non-green lineage). In the nongreen lineage, the
haptophytes formed a sister group to the clade containing heterokont algae,
red algae, and the cryptophyte Guillardia theta. This branching pattern was
well supported in terms of bootstrap values in weighted- parsimony and
maximum-likelihood analyses (100% and 92%, respectively). However, the
phylogenetic relationship among red algae, heterokonts, and a cryptophyte
taxon was not especially well resolved. A four- cluster analysis was
performed to further explore the statistical significance of the
relationship between proteobacteria, red algae (including and excluding
Guillardia theta), haptophytes, and heterokont algae. This test strongly
favored the hypothesis that the heterokonts and red algae are more closely
related to each other than either is to proteobacteria or haptophytes.
Hence, this molecular study based on a plastid-encoded gene provides
additional evidence for a distant relationship between haptophytes and the
heterokont algae. It suggests an evolutionary scenario in which the
ancestor of the haptophyte lineage engulfed a phototrophic eukaryote and,
more recently, the heterokont lineage became phototrophic by engulfing a
red alga.
相似文献