Shome Nath Mitra

Structure of the 1:1 netropsin–decamer d(CCIICICCII)2 complex with a single bound netropsin

The crystal structure of the 1:1 complex of netropsin and the B-DNA decamer d(CCIICICCII)(2) has been elucidated and refined to an R factor of 19.6% and an R(free) of 24.7% using 1790 reflections in the resolution range 8-2.4 A. The complex crystallizes in space group C2, with unit-cell parameters a = 62.40, b = 24.47, c = 36.31 A, beta = 110.09 degrees and one molecule of netropsin in the asymmetric unit; the rest of the minor groove is filled with six water molecules. The structure was solved by the molecular-replacement method using the DNA model d(CCCCCIIIII)(2) from the 2:1 netropsin complex by removing both bound netropsins (Chen et al., 1998). Surprisingly, only one netropsin molecule is found to bind to the present decamer, covering residues 2-6 at the upper stream of the duplex. The positively charged guanidinium head is hydrogen bonded through N1H(2) to the O(2) of cytosine 2 and through N10H(2) to N(3) of inosine 6. The three amide N-H groups of the peptides face the minor groove and form three sets of bifurcated hydrogen bonds with the base atoms. The central part of the drug (C3-N8) is nearly conjugated. The preference of the cytosine carbonyl O2 atoms over the inosine N3 atoms in hydrogen bonding is seen. The drug-bound region has more uniform twists, roll angles, propeller twists and minor-groove widths compared with the water-bound region.

Crystal Structure of an RNA Duplex [r(gugcaca)dC]2 with 3′-Dinucleoside Overhangs Forming a Superhelix

Abstract Crystal structure of the RNA octamer duplex, [r(gugcaca)dC] 2, with space group I212121 and the cell constants a=24.29, b=45.25 and c=73.68Å, has been determined and refined. The structural and packing architecture of the molecule consist of a highly bent six base paired duplex forming a right-handed superhelix stacked in tandem compared to an infinite pseudo- continuous column as is usually present in RNA crystal structures. The super helix could be formed by the head-to-head stacking (g1 over g1 and g9 over g9), the large bend and the twists at the junctions may also be responsible. The sugar-phosphate backbones of the 3′-end dinucleoside overhangs snuggly fit into the minor grooves of adjacent double helical stacks. The 3′-terminal deoxycytidines form antiparallel hemiprotonated trans (C·C)+ pairs with symmetry related deoxycytidines, while the penultimate adenines form base triples (a*g·c) with the capping g·c base pairs of the hexamer duplex with the adenine (a7) at one end being syn and at the other anti. These triple interactions are the same as those found in the tetrahymena ribozyme and group I intron.