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The Scaffolding Protein WAVE1 Associates with Kinesin 1 through the Tetratricopeptide Repeat (TPR) Domain of the Kinesin Light Chain (KLC) (Kinesin Light Chain (KLC)의 Tetratricopeptide Repeat (TPR) 도메인을 통한 Scaffold 단백질 WAVE1과 Kinesin 1의 결합)

  • Jang, Won Hee;Jeong, Young Joo;Urm, Sang-Hwa;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.8
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    • pp.963-969
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    • 2016
  • Kinesin superfamily proteins (KIFs) are microtubule-dependent molecular motor proteins essential for the intracellular transport of organelles and protein complexes in cells. Kinesin 1 is a member of those KIFs that transport various cargoes, including organelles, synaptic vesicles, neurotransmitter receptors, cell signaling molecules, and mRNAs through interaction between its light chain subunit and the cargoes. Kinesin light chains (KLCs) are non-motor subunits that associate with the kinesin heavy chain (KHC) dimer. KLCs interact with many different binding proteins, but their particular binding proteins have not yet been fully identified. We used the yeast two-hybrid assay to identify proteins that interact with the tetratricopeptide repeat (TPR) domain of KLC1. We found an interaction between the TPR domain of KLC1 and Wiskott-Aldrich syndrome protein family member 1 (WAVE1), a member of the WASP/WAVE family involved in regulation of actin cytoskeleton. WAVE1 bound to the six TPR domain-containing regions of KLC1 and did not interact with KHCs (KIF5A, KIF5B, and KIF5C) in the yeast two-hybrid assay. The carboxyl (C)-terminal verprolin-cofilin-acidic (VCA) domain of WAVE1 is essential for interaction with KLC1. Also, other WAVE isoforms (WAVE2 and WAVE3) interacted with KLC1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, WAVE1 co-localized with KLC1 and co-immunoprecipitated with KLC1 and KIF5B. These results suggest that kinesin 1 motor protein may transport WAVE complexes or WAVE-coated cargoes in cells.

APP Tail 1 (PAT1) Interacts with Kinesin Light Chains (KLCs) through the Tetratricopeptide Repeat (TPR) Domain (APP tail 1 (PAT1)과 kinesin light chains (KLCs)의 tetratricopeptide repeat (TPR) domain을 통한 결합)

  • Jang, Won Hee;Kim, Sang-Jin;Jeong, Young Joo;Jun, Hee Jae;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1608-1613
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    • 2012
  • A conventional kinesin, KIF5/Kinesin-I, transports various cargoes along the microtubule through interaction between its light chain subunit and the cargoes. Kinesin light chains (KLCs) interact with many different cargoes using their tetratricopeptide repeat (TPR) domain, but the mechanism underlying recognition and binding of a specific cargo has not yet been completely elucidated. We used the yeast two-hybrid assay to identify proteins that interact with the TPR domain of KLC1. We found an interaction between the TPR domain of KLC1 and an amyloid precursor protein (APP)-binding protein PAT1 (protein interacting with APP tail 1). The yeast two-hybrid assay demonstrated that the TPR domain-containing region of KLC1 mediated binding to the C-terminal tail region of PAT1. PAT1 also bound to KLC2 but not to kinesin heavy chains (KIF5A, KIF5B, and KIF5C) in the yeast two-hybrid assay. These protein-protein interactions were also observed in the glutathione S-transferase (GST) pull-down assay and by co-immunoprecipitation. Anti-PAT1 antibody as well as anti-APP anti-body co-immunoprecipitated KLC and KHCs associated with PAT1 from mouse brain extracts. These results suggest that PAT1 could mediate interactions between Kinesin-I and APP containing vesicles.

Analysis of Contamination Sources of Staphylococcus aureus Related to Perilla Leaves Using Multi-Locus Variable Number of Tandem Repeat Analysis (MLVA) (Multi-Locus Variable Number of Tandem Repeat Analysis (MLVA)를 이용한 들깻잎 중 Staphylococcus aureus의 오염원분석)

  • Kim, Se-Ri;Shim, Won-Bo;Han, Noo Ri;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.278-284
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    • 2014
  • To investigate the prevalence of Staphylococcus aureus (S. aureus) during production of perilla leaves, a total of 261 samples, including water, soil, surroundings of cultivation and packing, workers, and perilla leaves, was examined.. To trace the contamination sources of S. aureus related to perilla leaves, MLVA (Multi-Locous Variable number of tandem repeat Analysis), which is a very efficient method to discriminate strains with minimum molecular biology equipment was applied to S. aureus isolated from perilla leaves farms. S. aureus was isolated in perilla leaves from 9 of 38 farms at 0-2.92 log CFU/g. S. aureus was also found in working environment, including packing vinyl, worker clothes, irrigation water and hands. The patterns of MLVA of isolates from perilla leaves matched with those of isolates from packing table, irrigation water, packing vinyl, and hands. The isolates were successfully examined and determined by MLVA, thus elucidating S. aureus source and spread.

Effect of rbST Administrations at Artificial Insemination on Conception and Parturition Rates in Hanwoo (한우 인공수정시 rbST 투여가 수태 및 분만율에 미치는 영향)

  • Han M. H.;Choi S. H.;Choi Y. H.;Kim H. J.;Cho S. R.;Choi C.Y.;Ryu I. S.;Son D. S.;Yeon S. H.;Woo J. S.;Kweon U. G.;Yoon K. Y.;Chang B. S.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.177-184
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    • 2005
  • This study was conducted to investigate the effects of recombinant bovine somatotropin (rbST) injection on conception and parturition rates in normal or repeat breeding Hanwoo. We treated 462 cows containing 79 repeat-breeding cows of multiparous and allocating 5 treatment groups. Treatment 1 (T1) was injection of 2ml saline (for pseudo treatment), T2 was one injection of rbST 250mg into the tailhead region at the estrus, T3 was twice injection of rbST 250mg both at the time of insemination and again 10 to 14 day later, T4 was once injection of rbST 500mg at insemination and T5 was twice injection of 500mg rbST both at the time of insemination and again 10 to 14day later respectively. In rbST treated groups, timed artificial inseminations (TAI) were performed fellowing estrus synchronization. 100 us GnRH was injected into the scapula region on Day 0, 25mg $PGF_2{\alpha}$ was injected on Day 7 for degeneration of corpus luteum (CL) and 100ug GnRH was injected for inducing the synchronization. The results are as fellows; When normal Hanwoo were inseminated once with rbST administration, the pregnancy rate of T2 $(67.5\pm18.48\%)$ were higher than control $(52.4\pm9.72\%)$, while the pregnancy rate of T4 $(63.3\pm5.77\%)$ were significantly higher (p.<0.05) than control $(39.3\pm12.89\%)$ in repeat breeder Hanwoo. The parturition rates of normal Hanwoo were no differences among the treatments but were significant different in repeat breeder Hanwoo (p<0.05). When the estrous was induced by Ovsynch and inseminated once with rbST administration, the pregnancy rates of T2 was $12.5\%$ higher than control in normal Hanwoo, T4 $(80.0\%)$ was highest among the treatments (p<0.05) in repeat breeder Hanwoo. When normal Hanwoo were inseminated once with rbST administration, the pregnant period was $282.7\~284.8$ days and the body weight was $25.1\~25.9kg$, there were no difference among the treatments. The ratio of sex was almost same without T4 (male vs. female=18:9). In repeat breeder Hanwoo, pregnant period was 280.4~289.3 day and body weight was $23.0\~26.6kg$, it had no difference among the treatments. The sex ratio were similar to normal Hanwoo except T4 (M : F=2 : 8). In conclusion, the pregnancy and parturition rate by once insemination could be improved by the administration of rbST 250mg in normal Hanwoo or 500mg in repeat breeder Hawoo.

The Power and Pitfalls of Data Prefetching (데이터 미리읽기의 동작과 문제점)

  • Ki, An-do
    • Electronics and Telecommunications Trends
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    • v.13 no.4 s.52
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    • pp.59-69
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    • 1998
  • The terminology of data prefetching is introduced, which includes stride, repeat distance, stall, pending stall, prefetch degree, prefetch distance, and prefetch offset. The effectiveness of hardware data prefetching in reducing cache misses is shown by presenting a square matrix multiplication example. Thereafter the pitfalls of prefetching and possible solutions are discussed.