Template preference of polymerases and its relevance to oncogenic RNA virus replication

Zusammenfassung DNS-Polymerae aus Kalbsthymus (in50%-Glycerin-haltigem Puffer gelöst) zeigt bei Kühlaufbewahrung Alterung mit Änderung ihrer Template-Präferenz von einstrangiger zu doppelstrangiger DNS.

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Supported by U.S. Public Health Service and Damon Runyon Grants.     

Disordered RNA chaperone proteins: from functions to disease

RNA chaperones are ubiquitous proteins that play pivotal roles in cellular RNA metabolism and RNA virus replication. Here we propose that they act by organizing complex and highly dynamic networks of RNA-RNA, RNA-protein and protein-protein interactions. How this is achieved and how their malfunction may lead to disease will be discussed through the examples of human immunodeficiency virus type 1 nucleocapsid protein (NCp7), the fragile X mental retardation protein and the prion protein.Received 9 March 2005; received after revision 6 April 2005; accepted 6 April 2005Dedicated to the memory of Dominique Dormont     

RNA processing and human disease

Gene expression involves multiple regulated steps leading from gene to active protein. Many of these steps involve some aspect of RNA processing. Diseases caused by mutations that directly affect RNA processing are relatively rare compared with mutations that disrupt protein function. The vast majority of diseases of RNA processing result from loss of function of a single gene due to mutations in cis-acting elements required for pre-messenger RNA (mRNA) splicing. However, a few diseases are caused by alterations in the trans-acting factors required for RNA processing and in the vast majority of cases it is the pre-mRNA splicing machinery that is affected. Clearly, alterations that disrupt splicing of pre-mRNAs from large numbers of genes would be lethal at the cellular level. A common theme among these diseases is that only subsets of genes are affected. This is consistent with an emerging view that different subsets of exons require different sets of cis-acting elements and trans-acting factors.     

The potassium channel KcsA and its interaction with the lipid bilayer

The crystal structure of the K⁺ channel KcsA explains many features of ion channel function. The selectivity filter corresponds to a narrow region about 12 Å long and 3 Å wide, lined by carbonyl groups of the peptide backbone, through which a K⁺ ion can only move in a dehydrated form. The selectivity filter opens into a central, water-filled cavity leading to a gating site on the intracellular side of the channel. The channel is tetrameric, each monomer containing two transmembrane a helices, M1 and M2. Helix M1 faces the lipid bilayer and helix M2 faces the central channel pore; the M2 helices participate in subunit-subunit interactions. Helices M1 and M2 in each subunit pack as a pair of antiparallel coils with a heptad repeat, but the M2 helices of neighbouring subunits show fewer interactions, crossing at an angle of about –40°. Trp residues at the ends of the transmembrane helices form clear girdles on the two faces of the membrane, which, together with girdles of charged residues, define a hydrophobic thickness of about 37 Å for the channel. Binding constants for phosphatidylcholines to KcsA vary with fatty acyl chain length, the optimum chain length being C22. A phosphatidylcholine with this chain length gives a bilayer of thickness about 34 Å in the liquid crystalline phase, matching the hydrophobic thickness of the protein. However, a typical biological membrane has a hydrophobic thickness of about 27 Å. Thus either the transmembrane a helices of KcsA are more tilted in the native membrane than they are in the crystal structure, or the channel is under stress in the native membrane. The efficiency of hydrophobic matching between KcsA and the surrounding lipid bilayer is high over the chain length range C10–C24. The channel requires the presence of some anionic lipids for function, and fluorescence quenching studies show the presence of two classes of lipid binding site on KcsA; at one class of site (nonannular sites) anionic phospholipids bind more strongly than phosphatidylcholine, whereas at the other class of site (annular sites) phosphatidylcholines and anionic phospholipids bind with equal affinity.     

Presence of autocomplementary RNA with viral specificity in cells infected with herpes virus]

RNA from cells infected with Herpes simplex virus contain a higher percentage of double-stranded RNA than non-infected cells. This percentage increases three-fold upon self-annealing. The complementary RNA sequences were shown to be virus-specific by the following criteria: (1) high melting temperature than double-stranded RNA from non infected cells; (2) higher density in caesium sulphate; (3) specific hybridization with viral DNA.     

Epstein-barr virus and inflammatory bowel disease

Zusammenfassung Die Häufigkeit und der Antikörpertiter gegen Epstein-Barr-Virus (EBV) wurde in Patienten mit Ulkus-Colitis und Crohn's Krankheit, eine granulomatische Abnormalität ähnlich zur Sarkoidose untersucht und normale Häufigkeit und Antikörpertiter gegen EBV bei ihnen gefunden.

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Supported by Research Contract No. NIH-69-2078 within the Special Virus Cancer Program of the National Cancer Institute, National Institutes of Health, USPHS, National Institutes of Health Grant No. AM 05126 and the National Foundation for Ileitis and Colitis, Inc., New York, New York.

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Dr.Grotsky is a recipient of a National Institutes of Health Graduate Training Grant in Gastroenterology, No. Am 05126.

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Dr.Hirshaut is a recipient of a National Cancer Institute Grant No. CA 08748 an a grant from the New York Cancer Research Institute.

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Dr.Glade is a recipient of a Research Career Development Award No. A1-46371 of the USPHS.