Flap Endonuclease Disengages Dna2 Helicase/Nuclease from Okazaki Fragment Flaps

Okazaki fragments contain an initiator RNA/DNA primer that must be removed before the fragments are joined. In eukaryotes, the primer region is raised into a flap by the strand displacement activity of DNA polymerase {delta}. The Dna2 helicase/nuclease and then flap endonuclease 1 (FEN1) are proposed to act sequentially in flap removal. Dna2 and FEN1 both employ a tracking mechanism to enter the flap 5' end and move toward the base for cleavage. In the current model, Dna2 must enter first, but FEN1 makes the final cut at the flap base, raising the issue of how FEN1 passes the Dna2. To address this, nuclease-inactive Dna2 was incubated with a DNA flap substrate and found to bind with high affinity. FEN1 was then added, and surprisingly, there was little inhibition of FEN1 cleavage activity. FEN1 was later shown, by gel shift analysis, to remove the wild type Dna2 from the flap. RNA can be cleaved by FEN1 but not by Dna2. Pre-bound wild type Dna2 was shown to bind an RNA flap but not inhibit subsequent FEN1 cleavage. These results indicate that there is a novel interaction between the two proteins in which FEN1 disengages the Dna2 tracking mechanism. This interaction is consistent with the idea that the two proteins have evolved a special ability to cooperate in Okazaki fragment processing

General Rotorcraft Aeromechanical Stability Program (GRASP) version 1.03: User's manual

The Rotorcraft Dynamics Division, Aeroflightdynamics Directorate, U.S. Army Aviation Research and Technology Activity has developed the General Rotorcraft Aeromechanical Stability Program (GRASP) to perform calculations that will assess the stability of rotorcraft in hovering flight and ground contact conditions. The program is designed to be state-of-the-art, hybrid, finite-element/multibody code that can be applied to all existing and future helicopter configurations. While GRASP was specifically designed to solve rotorcraft stability problems, its innovative structure and formulation allow for application to a wide range of structures. This manual describes the preparation of the input file required by Version 1.03 of GRASP, the procedures used to invoke GRASP on the NASA Ames Research Center CRAY X-MP 48 computer, and the interpretation of the output produced by GRASP. The parameters used by the input file are defined, and summaries of the input file and the job control language are included

Catalytic surface effects on contaminated space shuttle tile in a dissociated nitrogen stream

Visual inspection revealed contamination on the surface of tiles removed from the lower section of the space shuttle orbiter after the second flight of Columbia (STS-2). Possible sources of this contamination and the effect on surface catalycity are presented

Supercharged topping rocket propellant feed system

A rocket propellant feed system utilizing a bleed turbopump to supercharge a topping turbopump is presented. The bleed turbopump is of a low pressure type to meet the cavitation requirements imposed by the propellant storage tanks. The topping turbopump is of a high pressure type and develops 60 to 70 percent of the pressure rise in the propellant

Use of similarity parameters for examination of geometry characteristics of high-expansion- ratio axial-flow turbines

Similarity parameters used for examining geometry characteristics of axial flow turbines with high expansion rati

COLLECTIVE ACTION AND INFORMAL FINANCIAL INSTITUTIONS: AN EMPIRICAL ANALYSIS OF ROTATING AND SAVINGS CREDIT ASSOCIATIONS (ROSCAS) IN SENEGAL

This study analyzes how rotating savings and credit associations (ROSCAs) in Senegal were able to overcome the collective action dilemma, maintain institutional performance, and remain sustainable over time. This study models cooperation among members as well as the performance and sustainability of associations using data collected from field research conducted in Dakar, Senegal in 2001. The results show that factors such as homogeneity of individuals within an association, how long the association has existed, how defaults are covered, and rules such residency requirements, individual contributions, and rotation order are to various degree critical to the performance and sustainability of ROSCAs and to the fostering of cooperation among members of these associations.Financial Economics,

The application of experimental data to blade wake interaction noise prediction

Blade wake interaction noise (BWI) has been defined as the broadband noise generated by the ingestion of turbulent trailing tip vortices by helicopter rotors. This has been shown to be the dominant contributor to the subjectively important part of the acoustic spectrum for the approach stage of a helicopter flyover. A prediction method for BWI noise based on the calculated trailing vortex trajectories has been developed and estimates of the vortex turbulence have been made. These measurements were made on a trailing vortex from a split wing arrangement and did not give the spectrum of the velocity fluctuations. A recent experiment carried out to measure the turbulence associated with a trailing vortex and the application of the results to BWI noise prediction is described

The Political and Economic Determinants of Trade Disputes under the WTO

Replaced with revised version of paper 02/25/08.democratization, system of government, trade disputes, World Trade Organization (WTO), economic strata, International Relations/Trade, P16, F13,

Dynamic removal of replication protein A by Dna2 facilitates primer cleavage during Okazaki fragment processing in Saccharomyces cerevisiae

Eukaryotic Okazaki fragments are initiated by an RNA/DNA primer, which is removed before the fragments are joined. Polymerase d displaces the primer into a flap for processing. Dna2 nuclease/helicase and flap endonuclease 1 (FEN1) are proposed to cleave the flap. The single-stranded DNA binding protein, replication protein A (RPA), governs cleavage activity. Flap-bound RPA inhibits FEN1. This necessitates cleavage by Dna2, which is stimulated by RPA. FEN1 then cuts the remaining RPA-free flap to create a nick for ligation. Cleavage by Dna2 requires that it enter the 5'-end and track down the flap. Since Dna2 cleaves the RPA-bound flap, we investigated the mechanism by which Dna2 accesses the protein-coated flap for cleavage. Using a nuclease-defective Dna2 mutant, we showed that just binding of Dna2 dissociates the flap-bound RPA. Facile dissociation is specific to substrates with a genuine flap, and will not occur with an RPA-coated single strand. We also compared the cleavage patterns of Dna2 with and without RPA to better define RPA stimulation of Dna2. Stimulation derived from removal of DNA folding in the flap. Apparently, coordinated with its dissociation, RPA relinquishes the flap to Dna2 for tracking in a way that does not allow flap structure to reform. We also found that RPA strand melting activity promotes excessive flap elongation, but it is suppressed by Dna2-promoted RPA dissociation. Overall, results indicate that Dna2 and RPA coordinate their functions for efficient flap cleavage and preparation for FEN1