Toward Monodisperse Poly(γ-benzyl α,L-glutamate): Uniform, Polar, Molecular Rods

Poly(γ-benzyl α,L-glutamate) (PBLG) has been widely used in studies of the physics of rod-like polymer chains. The helical structure of PBLG gives rise to considerable chain stiffness, such that the persistence length of the chain is on the order of 70 nm in helicogenic solvents. This feature, coupled with the ease of synthesis and good solubility of the polymer has made PBLG the system of choice for the study of both isotropic and liquid crystalline solutions of rod-like macromolecules

Comments on the Role of Molecular Genetics in Polymer Materials Science

The most fundamental goal of the synthetic chemist is control of molecular architecture. With respect to small molecules (i.e., those of molecular weight less than a few thousand), this means absolute control of chemical connectivity and stereochemistry – complete specification of molecular structure. But in macromolecular chemistry, controlled architecture has meant something quite different. Because polymerizations are in general statistical processes, conventional polymeric materials are characterized by substantial heterogeneity in chain length, sequence and stereochemistry. Control is exercised in a statistical sense only, and considerable skill is required to control even the average properties of the chain population and the dispersity in those properties

Synthesis and Characterization of Periodic Polypeptides Containing Repeating —(AlaGly)_xGluGly— Sequences

We have expressed in E. coli a series of periodic polypeptides represented by sequence 1. Our objective has been an understanding of the role of chemical sequence in determining the chain folding behavior of periodic macromolecules. Molecular organization has been examined by infrared spectroscopy and ^1H and ^(13)C NMR methods and a preliminary model of the folded structure has been developed

The Helicase Has1p Is Required for snoRNA Release from Pre-rRNA

Synthesis of rRNA in eukaryotes involves the action of a large population of snoRNA-protein complexes (snoRNPs), which create modified nucleotides and participate in cleavage of pre-rRNA. The snoRNPs mediate these functions through direct base pairing, in many cases through long complementary sequences. This feature suggests that RNA helicases may be involved in the binding and release of snoRNPs from pre-rRNA. In this study, we determined that the DEAD box helicase Has1p, a nucleolar protein required for the production of 18S rRNA, copurifies with the snR30/U17 processing snoRNP but is also present with other snoRNPs. Blocking Has1p expression causes a substantial increase in snoRNPs associated with 60S-90S preribosomal RNP complexes, including the U3 and U14 processing snoRNPs and several modifying snoRNPs examined. Cosedimentation persisted even after deproteinization. This effect was not observed with depletion of two nonhelicase proteins, Esf1p and Dim2p, that are also required for 18S rRNA production. Point mutations in ATPase and helicase motifs of Has1p block U14 release from pre-rRNA. Surprisingly, depletion of Has1p causes a reduction in the level of free U6 snRNP. The results indicate that the Has1p helicase is required for snoRNA release from pre-rRNA and production of the U6 snRNP

Interference probing of rRNA with snoRNPs: A novel approach for functional mapping of RNA in vivo

Synthesis of eukaryotic ribosomal RNAs (rRNAs) includes methylation of scores of nucleotides at the 2′-O-ribose position (Nm) by small nucleolar RNP complexes (snoRNPs). Sequence specificity is provided by the snoRNA component through base-pairing of a guide sequence with rRNA. Here, we report that methylation snoRNPs can be targeted to many new sites in yeast rRNA, by providing the snoRNA with a novel guide sequence, and that in some cases growth and translation activity are strongly impaired. Novel snoRNAs can be expressed individually or by a unique library strategy that yields guide sequences specific for a large target region. Interference effects were observed for sites in both the small and large subunits, including the reaction center region. Targeting guide RNAs to nucleotides flanking the sensitive sites caused little or no defect, indicating that methylation is responsible for the interference rather than a simple antisense effect or misguided chaperone function. To our knowledge, this is the only approach that has been used to mutagenize the backbone of rRNA in vivo

Functional Mapping of the U3 Small Nucleolar RNA from the Yeast Saccharomyces cerevisiae

The U3 small nucleolar RNA participates in early events of eukaryotic pre-rRNA cleavage and is essential for formation of 18S rRNA. Using an in vivo system, we have developed a functional map of the U3 small nucleolar RNA from Saccharomyces cerevisiae. The test strain features a galactose-dependent U3 gene in the chromosome and a plasmid-encoded allele with a unique hybridization tag. Effects of mutations on U3 production were analyzed by evaluating RNA levels in cells grown on galactose medium, and effects on U3 function were assessed by growing cells on glucose medium. The major findings are as follows: (i) boxes C � and D and flanking helices are critical for U3 accumulation; (ii) boxes B and C are not essential for U3 production but are important for function, most likely due to binding of a trans-acting factor(s); (iii) the 5 � portion of U3 is required for function but not stability; and, (iv) strikingly, the nonconserved hairpins 2, 3, and 4, which account for 50 % of the molecule, are not required for accumulation or function. The small nucleolar RNAs (snoRNAs) play essential roles in posttranscriptional maturation of rRNAs (reviewed in reference