Combined transcriptome and proteome analysis as a powerful approach to study genes under glucose repression in Bacillus subtilis

We used 2D protein gel electrophoresis and DNA microarray technologies to systematically analyze genes under glucose repression in Bacillus subtilis. In particular, we focused on genes expressed after the shift from glycolytic to gluconeogenic at the middle logarithmic phase of growth in a nutrient sporulation medium, which remained repressed by the addition of glucose. We also examined whether or not glucose repression of these genes was mediated by CcpA, the catabolite control protein of this bacterium. The wild-type and ccpA1 cells were grown with and without glucose, and their proteomes and transcriptomes were compared. 2D gel electrophoresis allowed us to identify 11 proteins, the synthesis of which was under glucose repression. Of these proteins, the synthesis of four (IolA, I, S and PckA) was under CcpA-independent control. Microarray analysis enabled us to detect 66 glucose-repressive genes, 22 of which (glmS, acoA, C, yisS, speD, gapB, pckA, yvdR, yxeF, iolA, B, C, D, E, F, G, H, I, J, R, S and yxbF ) were at least partially under CcpA-independent control. Furthermore, we found that CcpA and IolR, a repressor of the iol divergon, were involved in the glucose repression of the synthesis of inositol dehydrogenase encoded by iolG included in the above list. The CcpA-independent glucose repression of the iol genes appeared to be explained by inducer exclusion.

GenEST, a powerful bidirectional link between cDNA sequence data and gene expression profiles generated by cDNA-AFLP

The release of vast quantities of DNA sequence data by large-scale genome and expressed sequence tag (EST) projects underlines the necessity for the development of efficient and inexpensive ways to link sequence databases with temporal and spatial expression profiles. Here we demonstrate the power of linking cDNA sequence data (including EST sequences) with transcript profiles revealed by cDNA-AFLP, a highly reproducible differential display method based on restriction enzyme digests and selective amplification under high stringency conditions. We have developed a computer program (GenEST) that predicts the sizes of virtual transcript-derived fragments (TDFs) of in silico-digested cDNA sequences retrieved from databases. The vast majority of the resulting virtual TDFs could be traced back among the thousands of TDFs displayed on cDNA-AFLP gels. Sequencing of the corresponding bands excised from cDNA-AFLP gels revealed no inconsistencies. As a consequence, cDNA sequence databases can be screened very efficiently to identify genes with relevant expression profiles. The other way round, it is possible to switch from cDNA-AFLP gels to sequences in the databases. Using the restriction enzyme recognition sites, the primer extensions and the estimated TDF size as identifiers, the DNA sequence(s) corresponding to a TDF with an interesting expression pattern can be identified. In this paper we show examples in both directions by analyzing the plant parasitic nematode Globodera rostochiensis. Various novel pathogenicity factors were identified by combining ESTs from the infective stage juveniles with expression profiles of ∼4000 genes in five developmental stages produced by cDNA-AFLP.

Immucillin H, a powerful transition-state analog inhibitor of purine nucleoside phosphorylase, selectively inhibits human T lymphocytes

Transition-state theory has led to the design of Immucillin-H (Imm-H), a picomolar inhibitor of purine nucleoside phosphorylase (PNP). In humans, PNP is the only route for degradation of deoxyguanosine, and genetic deficiency of this enzyme leads to profound T cell-mediated immunosuppression. This study reports the biological effects and mechanism of action of Imm-H on malignant T cell lines and on normal activated human peripheral T cells. Imm-H inhibits the growth of malignant T cell leukemia lines with the induction of apoptosis. Imm-H also inhibits activated normal human T cells after antigenic stimulation in vitro. However, Imm-H did not inhibit malignant B cells, colon cancer cell lines, or normal human nonstimulated T cells, demonstrating the selective activity of Imm-H. The effects on leukemia cells were mediated by the cellular phosphorylation of deoxyguanosine and the accumulation of dGTP, an inhibitor of ribonucleotide diphosphate reductase. Cells were protected from the toxic effects of Imm-H when deoxyguanosine was absent or when deoxycytidine was present. Guanosine incorporation into nucleic acids was selectively blocked by Imm-H with no effect on guanine, adenine, adenosine, or deoxycytidine incorporation. Imm-H may have clinical potential for treatment of human T cell leukemia and lymphoma and for other diseases characterized by abnormal activation of T lymphocytes. The design of Imm-H from an enzymatic transition-state analysis exemplifies a powerful approach for developing high-affinity enzyme inhibitors with pharmacologic activity.

Evolution of powerful extragalactic radio sources.

Observations of complete flux density limited samples of powerful extragalactic radio sources by very-long-baseline interferometry enable us to study the evolution of these objects over the range of linear scales from 1 parsec to 15 kiloparsees (1 parsec = 3.09 x 10(18) cm). The observations are consistent with the unifying hypothesis that compact symmetric objects evolve into compact steep-spectrum doubles, which in turn evolve into large-scale Fanaroff-Riley class II objects. It is suggested that this is the primary evolutionary track of powerful extragalactic radio sources. In this case there must be significant luminosity evolution in these objects, but little velocity evolution, as they expand from 1 parsec to several hundred kiloparsecs in overall size.

The most powerful man in the world; the life of Sir Henri Deterding.

Mode of access: Internet.

A key to species of American owls,

Mode of access: Internet.

The versenes, powerful organic chelating agents for the exacting chemical control of cations.

Mode of access: Internet.

Powerful ideas-- : --that saves Army energy.

Mode of access: Internet.

The History and romance of ancient empires; the rise to power, the conquest, dominion and downfall of the powerful nations of antiquity.

Mode of access: Internet.

A cursory view of the Creation; in a hymn to the all-gracious, wise and powerful Creator. In four parts.

Unbound, in cloth case.

Saugu Asmundar, er Kalladur er Kappabani : eller Asmunds Kappabanes saga, hoc est narratio historica rerum præclare gestarum, ab Asmundo, cui strenua dextra cognomen, pugilum interfectoris peperit; exhibens insuper acta regum, uti sunt: budlii svioniæ, alfiique Daniæ; nec non hildibrandorum, hunalandijæ olim ducem ac heroum, martia facinora commemorans. : hanc, e cod. Mscto. membranaceo, linguæ priscæ Scandicæ, descriptam, cum gemina versione /

Parallel text in Latin, Danish, and Old Norse (?).

Rescued from fiery death; a powerful narrative of the Iroquois theater disaster,

Mode of access: Internet.

Conversations between two laymen on strict and mixed communion : in which the principal arguments in favor of the latter practice, are stated, as nearly as possible, in the words of its most powerful advocate, the Rev. Robert Hall /

Title on half t.p.: Views of communion / by Hall, Fuller, Griffin and Ripley.