38 research outputs found

    Single molecule fluorescence studies of the RNA polymerase II elongation complex

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    Peranan Kepemimpinan Camat Dalam Menumbuhkan Kedisiplinan Pegawai Di Kantor Kecamatan Jebres Kota Surakarta

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    Absract: The objectives of this research are to investigate: (1) how Chief of Jebres Sub-district Office, Surakarta city plays his leadership role in growing the discipline behavior of its employees; (2) what constraints are encountered by Chief of Jebres Sub-district Office, Surakarta city in internalizing the discipline of its employees; and (3) what solutions are taken by Chief of Jebres Sub-district Office to deal with the prevailing constraints to the materialization of the discipline behavior of its employees. This research used the qualitative embedded single case study method. The results of the research are as follows: 1) Chief of Jebres Sub-district Office, Surakarta city plays his leadership role through the discipline behavior internalizations such as: (a) giving exemplification through discipline attitude; (b) giving motivation to the employees; (c) attempting to fulfill their needs and prosperity; and (d) implementing the discipline upholding consistently. 2) The constraints occurring in nurturing the discipline of the employees of Jebres Sub-district Office are as follows: (a) internal constraints: some employees bear the characters and personalities which are apathetic to advices, and some are lack of responsibilities in executing their duties and work and (b) external constraints: the external constraints occurring in internalizing the discipline of the employees of Jebres Sub-district Office are mainly related to the work environment. 3) The prevailing constraints occurring in internalizing the discipline of the employees are as follows: (a) upholding the prevailing laws and regulations and nurturing the employees; and (b) maintaining the communication and good relations among the employees

    Nano positioning system reveals the course of upstream and nontemplate DNA within the RNA polymerase II elongation complex

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    Crystallographic studies of the RNA polymerase II (Pol II) elongation complex (EC) revealed the locations of downstream DNA and the DNA-RNA hybrid, but not the course of the nontemplate DNA strand in the transcription bubble and the upstream DNA duplex. Here we used single-molecule Fluorescence Resonance Energy Transfer (smFRET) experiments to locate nontemplate and upstream DNA with our recently developed Nano Positioning System (NPS). In the resulting complete model of the Pol II EC, separation of the nontemplate from the template strand at position +2 involves interaction with fork loop 2. The nontemplate strand passes loop β10-β11 on the Pol II lobe, and then turns to the other side of the cleft above the rudder. The upstream DNA duplex exits at an approximately right angle from the incoming downstream DNA, and emanates from the cleft between the protrusion and clamp. Comparison with published data suggests that the architecture of the complete EC is conserved from bacteria to eukaryotes and that upstream DNA is relocated during the initiation–elongation transition

    Direct observation and control of supported lipid bilayer formation with interferometric scattering microscopy

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    Supported lipid bilayers (SLB) are frequently used to study processes associated with or mediated by lipid membranes. The mechanism by which SLBs form is a matter of debate, largely due to the experimental difficulty associated with observing the adsorption and rupture of individual vesicles. Here, we used interferometric scattering microscopy (iSCAT) to directly visualize membrane formation from nanoscopic vesicles in real time. We observed a number of previously proposed phenomena such as vesicle adsorption, rupture, movement, and a wave-like bilayer spreading. By varying the vesicle size and the lipid-surface interaction strength, we rationalized and tuned the relative contributions of these phenomena to bilayer formation. Our results support a model where the interplay between bilayer edge tension and the overall interaction energy with the surface determine the mechanism of SLB formation. The unique combination of sensitivity, speed, and label-free imaging capability of iSCAT provides exciting prospects not only for investigations of SLB formation, but also for studies of assembly and disassembly processes on the nanoscale with previously unattainable accuracy and sensitivity
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