Environmental Pressure May Change the Composition Protein Disorder in Prokaryotes

Many prokaryotic organisms have adapted to incredibly extreme habitats. The genomes of such extremophiles differ from their non-extremophile relatives. For example, some proteins in thermophiles sustain high temperatures by being more compact than homologs in non-extremophiles. Conversely, some proteins have increased volumes to compensate for freezing effects in psychrophiles that survive in the cold. Here, we revealed that some differences in organisms surviving in extreme habitats correlate with a simple single feature, namely the fraction of proteins predicted to have long disordered regions. We predicted disorder with different methods for 46 completely sequenced organisms from diverse habitats and found a correlation between protein disorder and the extremity of the environment. More specifically, the overall percentage of proteins with long disordered regions tended to be more similar between organisms of similar habitats than between organisms of similar taxonomy. For example, predictions tended to detect substantially more proteins with long disordered regions in prokaryotic halophiles (survive high salt) than in their taxonomic neighbors. Another peculiar environment is that of high radiation survived, e.g. by Deinococcus radiodurans. The relatively high fraction of disorder predicted in this extremophile might provide a shield against mutations. Although our analysis fails to establish causation, the observed correlation between such a simplistic, coarse-grained, microscopic molecular feature (disorder content) and a macroscopic variable (habitat) remains stunning.     

Characterization of the central region containing the X-inactivation center and terminal region of the mouse X Chromosome using irradiation and fusion gene transfer hybrids

The irradiation and fusion gene transfer (IFGT) procedure provides a means of isolating subchromosomal fragments for use in the mapping of loci and for cloning probes from a particular area of a chromosome. Using this procedure, two large panels of somatic cell hybrids that contain mouse X Chromosome (Chr) fragments have been generated. These hybrid panels were generated by irradiating the monochromosomal mouse-hamster hybrid HYBX, which retains the mouse X Chr, with either 10 K or 50 K rads of X-irradiation followed by fusion with a recipient Chinese hamster cell line. IFGT hybrids retaining mouse material were generated at high frequency. These hybrids were used to orient loci in the X-inactivation center region that had not been resolvable in our interspecies backcross panel and also to map, within the terminal region of the X Chr, repeat elements detected by the probe p15-4. These hybrids not only complement existing interspecies meiotic mapping panels for the detailed analysis of specific regions of particular chromosomes, but also provide a potential source of material for chromosome-specific probe isolation.     

Genetic and molecular evidence of an X-chromosome deletion spanning the tabby (Ta) and testicular feminization (Tfm) loci in the mouse

A new radiation-induced mutation in the mouse, tabby-25H (Ta25H), has proved to be a deletion which spans both the tabby and testicular feminization (Tfm) loci on the X chromosome. The Ta phenotype closely resembles that of the original TaFa mutation in both the heterozygous and hemizygous conditions but Ta25H/Y animals additionally show the Tfm/Y phenotype, being externally female but possessing abdominally located testes. There is a shortage of both Ta25H/+ and Ta25H/Y classes relative to their normal sibs among the progeny of Ta25H/+ females at weaning age and this was indicated to be due to prenatal or neonatal losses. Exencephaly was observed in some members of both classes prior to birth. Both Ta25H classes tend to be runted at weaning but, remarkably, Ta25H/+ females often show a range of abnormalities not evident in Ta25H/Y animals. When probes for the Zfx, Ccg-1, Phk, and DXPas19 loci, which lie close to Ta, were hybridised to DNAs from Ta25H hemizygotes, the profiles of the X-linked bands were similar to those of control DNAs, suggesting these loci lie outside the deletion. However, a clear absence of an X-linked band was found with human androgen receptor probes, indicating that the Tfm locus is indeed missing. The deletion, therefore, extends a minimum of 1.5 cM and, with its proximal and distal boundaries partially defined, it could be as large as 4 cM. As Ta25H/+ females show the striped X-inactivation coat pattern, the putative X-inactivation centre, Xce, which lies close to Ta, cannot be located within the region deleted. The greasy (Gs) locus similarly appears to lie outside the deletion.     

Mapping of Murine Fibroblast Growth Factor Receptors Refines Regions of Homology between Mouse and Human Chromosomes

The genes for the fibroblast growth factor receptors Fgfr2, Fgfr3, and Fgfr4 have been mapped in the mouse using an interspecific backcross mapping panel. The Fgfr loci map to previously defined regions of homology between human and mouse chromosomes and provide additional information regarding human/mouse comparative mapping.     

Crystal structure of the collagen alpha1(VIII) NC1 trimer.

Collagen VIII is a major component of Descemet's membrane and is also found in vascular subendothelial matrices. The C-terminal non-collagenous domain (NC1) domain of collagen VIII, which is a member of the C1q-like protein family, forms a stable trimer and is thought to direct the assembly of the collagen triple helix, as well as polygonal supramolecular structures. We have solved the crystal structure of the mouse alpha1(VIII)(3) NC1 domain trimer at 1.9 A resolution. Each subunit of the intimate NC1 trimer consists of a ten-stranded beta-sandwich. The surface of the collagen VIII NC1 trimer presents three strips of partially exposed aromatic residues shown to interact with the non-ionic detergent CHAPS, which are likely to be involved in supramolecular assemblies. Equivalent strips exist in the NC1 domain of the closely related collagen X, suggesting a conserved assembly mechanism. Surprisingly, the collagen VIII NC1 trimer lacks the buried calcium cluster of the collagen X NC1 trimer. The mouse alpha1(VIII) and alpha2(VIII) NC1 domains are 71.5% identical in sequence, with the differences being concentrated on the NC1 trimer surface. A few non-conservative substitutions map to the subunit interfaces near the surface, but it is not obvious from the structure to what extent they determine the preferred assembly of collagen VIII alpha1 and alpha2 chains into homotrimers.     

Mouse X chromosome

Overall, the probe map fromDXWas70 toAmg encompasses 72 cM and includes 103 loci. Eight of these have been designated reference loci (see Table 2 and previous section) on account of their wide usage that would enable the cross reference of independent maps created in different laboratories. Reference loci are to be readily available and easily used probes for Southern hybridization. By and large, they will be STSs, regeneratable by PCR, and correspond to a known gene. In addition, on the mouse X Chr, there is a reference locus from each of the known conserved linkage groups found between the mouse and human X Chrs. All the loci, barDXWas31, represent conserved sequences. Committee Members: D. Adler, P.J. Barnard, Y. Boyd, N. Brockdorff, J. Derry, C. Disteche, C. Faust, M.F. Lyon, S. Rastan, M. Seldin and L. Siracusa.     

Processing of snake venom l-amino acid oxidase during intracellular transport

The isoelectrofocusing patterns of l-amino acid oxidase (LAO) from venom gland homogenates and of the secreted venom of Vipera palaestinae have been compared. The LAO isozyme profile of actively synthesizing gland spans over a wider range of pIs (4.8–6.0) and includes more variants as compared with the profile of the secreted venom. A basic shift of the isoelectrofocusing pattern of LAO obtained by treatment of the gland homogenate or the venom with neuraminidase indicates that sialic acid residues are responsible for the changes in the electronegativity of the isozymes. Analyses of subcellular fractions show that the microsomal fraction of the venom gland homogenate exhibits the highest multiplicity of molecular forms of LAO, whereas the fraction including the secretory granules has an isozyme profile similar to the venom. Double labelling experiments show that the newly synthesized LAO include isozymes which span over a wide range of pIs, whereas later on labelling of the more acidic isozymes is prominent. The results obtained may suggest that the sialic acid residues which are attached to LAO during its transport serve as “markers” for secretion.     

Distinctive properties of fucosyl glycopeptides on human teratoma cells

Fucose-labeled glycopeptides from four human teratoma cell lines of independent origin show similar elution profiles on Sephadex G-50 column chromatography. The fucosyl glycopeptides elute in two major regions: one near the void volume, the other in fractions corresponding to a molecular weight of 2500-3000. These elution profiles are very different from those obtained with the other human cell lines examined which included 3 lymphomas, 2 colon carcinomas, and HeLa. The elution profiles of the human teratomas, however, show remarkable similarities to those obtained with murine embryonal carcinoma cell culture and early mouse embryos. These results suggest that the excluded G-50 fraction may well contain glycopeptides playing a role in mammalian embryogenesis.