Regulation of Ack1 localization and activity by the amino-terminal SAM domain

<p>Abstract</p> <p>Background</p> <p>The mechanisms that regulate the activity of the nonreceptor tyrosine kinase Ack1 (activated Cdc42-associated kinase) are poorly understood. The amino-terminal region of Ack1 is predicted to contain a sterile alpha motif (SAM) domain. SAM domains share a common fold and mediate protein-protein interactions in a wide variety of proteins. Here, we addressed the importance of the Ack1 SAM domain in kinase activity.</p> <p>Results</p> <p>We used immunofluorescence and Western blotting to show that Ack1 deletion mutants lacking the N-terminus displayed significantly reduced autophosphorylation in cells. A minimal construct comprising the N-terminus and kinase domain (NKD) was autophosphorylated, while the kinase domain alone (KD) was not. When expressed in mammalian cells, NKD localized to the plasma membrane, while KD showed a more diffuse cytosolic localization. Co-immunoprecipitation experiments showed a stronger interaction between full length Ack1 and NKD than between full length Ack1 and KD, indicating that the N-terminus was important for Ack1 dimerization. Increasing the local concentration of purified Ack1 kinase domain at the surface of lipid vesicles stimulated autophosphorylation and catalytic activity, consistent with a requirement for dimerization and trans-phosphorylation for activity.</p> <p>Conclusions</p> <p>Collectively, the data suggest that the N-terminus of Ack1 promotes membrane localization and dimerization to allow for autophosphorylation.</p

The life and scientific work of William R. Evitt (1923-2009)

Occasionally (and fortunately), circumstances and timing combine to allow an individual, almost singlehandedly, to generate a paradigm shift in his or her chosen field of inquiry. William R. (‘Bill’) Evitt (1923-2009) was such a person. During his career as a palaeontologist, Bill Evitt made lasting and profound contributions to the study of both dinoflagellates and trilobites. He had a distinguished, long and varied career, researching first trilobites and techniques in palaeontology before moving on to marine palynomorphs. Bill is undoubtedly best known for his work on dinoflagellates, especially their resting cysts. He worked at three major US universities and spent a highly significant period in the oil industry. Bill's early profound interest in the natural sciences was actively encouraged both by his parents and at school. His alma mater was Johns Hopkins University where, commencing in 1940, he studied chemistry and geology as an undergraduate. He quickly developed a strong vocation in the earth sciences, and became fascinated by the fossiliferous Lower Palaeozoic strata of the northwestern United States. Bill commenced a PhD project on silicified Middle Ordovician trilobites from Virginia in 1943. His doctoral research was interrupted by military service during World War II; Bill served as an aerial photograph interpreter in China in 1944 and 1945, and received the Bronze Star for his excellent work. Upon demobilisation from the US Army Air Force, he resumed work on his PhD and was given significant teaching duties at Johns Hopkins, which he thoroughly enjoyed. He accepted his first professional position, as an instructor in sedimentary geology, at the University of Rochester in late 1948. Here Bill supervised his first two graduate students, and shared a great cameraderie with a highly motivated student body which largely comprised World War II veterans. At Rochester, Bill continued his trilobite research, and was the editor of the Journal of Paleontology between 1953 and 1956. Seeking a new challenge, he joined the Carter Oil Company in Tulsa, Oklahoma, during 1956. This brought about an irrevocable realignment of his research interests from trilobites to marine palynology. He undertook basic research on aquatic palynomorphs in a very well-resourced laboratory under the direction of one of his most influential mentors, William S. ‘Bill’ Hoffmeister. Bill Evitt visited the influential European palynologists Georges Deflandre and Alfred Eisenack during late 1959 and, while in Tulsa, first developed several groundbreaking hypotheses. He soon realised that the distinctive morphology of certain fossil dinoflagellates, notably the archaeopyle, meant that they represent the resting cyst stage of the life cycle. The archaeopyle clearly allows the excystment of the cell contents, and comprises one or more plate areas. Bill also concluded that spine-bearing palynomorphs, then called hystrichospheres, could be divided into two groups. The largely Palaeozoic spine-bearing palynomorphs are of uncertain biological affinity, and these were termed acritarchs. Moreover, he determined that unequivocal dinoflagellate cysts are all Mesozoic or younger, and that the fossil record of dinoflagellates is highly selective. Bill was always an academic at heart and he joined Stanford University in 1962, where he remained until retiring in 1988. Bill enjoyed getting back into teaching after his six years in industry. During his 26-year tenure at Stanford, Bill continued to revolutionise our understanding of dinoflagellate cysts. He produced many highly influential papers and two major textbooks. The highlights include defining the acritarchs and comprehensively documenting the archaeopyle, together with highly detailed work on the morphology of Nannoceratopsis and Palaeoperidinium pyrophorum using the scanning electron microscope. Bill supervised 11 graduate students while at Stanford University. He organised the Penrose Conference on Modern and Fossil Dinoflagellates in 1978, which was so successful that similar meetings have been held about every four years since that inaugural symposium. Bill also taught many short courses on dinoflagellate cysts aimed at the professional community. Unlike many eminent geologists, Bill actually retired from actively working in the earth sciences. His full retirement was in 1988; after this he worked on only a small number of dinoflagellate cyst projects, including an extensive paper on the genus Palaeoperidinium

On Application of Ring Linear Feedback Shift Registers to Testing of Interconnects in FPGAs

Praca poświęcona jest dedykowanemu konkretnej aplikacji testowaniu połączeń w układach FPGA. Na czas testowania komórki układu FPGA wchodzące w skład realizowanej aplikacji są przekształcane w elementy układu RL-BIST. Do budowy takiego układu został wybrany pierścieniowy rejestr LFSR, którego n pętli sprzężeń zwrotnych jest w trakcie testowania liniami testowanej magistrali połączeń. Na podstawie sygnatury otrzymanej w układzie RL-BIST stwierdza się czy testowana magistrala połączeń jest sprawna a w oparciu o słownik diagnostyczny można także zlokalizować uszkodzone połączenia oraz zidentyfikować typ uszkodzenia. Skuteczność zaproponowanej metody testowania połączeń w FPGA została poparta obszernymi wynikami eksperymentalnymi.Due to rapidly growing complexity of FPGA circuits application-dependent techniques of their testing become more and more often exploited for manufacturing test instead of application'independent methods. In such the case not all but only a part of FPGA resources (i.e. CLBs and interconnects) is a subject of testing - the part that is to be used by the concrete target application. The work is devoted to application-dependent testing of interconnects in FPGA circuits. For the test period the CLBs being the parts of the application are reconfigured so they implement elements (i.e. XOR gates and D-type flip-flops) of a RL-BIST structure based on a ring linear feedback shift register (R-LFSR). FPGA interconnections under test (IUTs) or at least their part are feedback lines of the R-LFSR. The R-LFSR is first initialised with a randomly chosen seed and than run for several clock cycles. Next the final state of the R-LFSR - a signature - is red by an ATE (Automatic Test Equipment). The value of the signature determines whether IUTs are fault free or faulty. Moreover, on the basis of the signature and with the use of a fault dictionary one may localise faulty interconnections in the FPGA and identify types of faults. The FPGA is afterwards reconfigured so the other set of IUTs becomes feedback lines of the R-LFSR. The above procedure is repeated until all FPGA interconnections belonging to the target application are tested. Efficacy of the proposed approach to testing of FPGA interconnects is supported by experimental results

On the use of a ring LFSR for testing crosstalk faults in interconnect networks

W pracy przedstawiono nową metodę wykrywania przesłuchów w połączeniach. Testowaniu poddaje się tylko te połączenia FPGA, które będą wykorzystywane przez docelową aplikację. Zaproponowana struktura testera wbudowanego (BIST) wykorzystuje rejestr pierścieniowy 3n R LFSR, który w swojej części odpowiedzialnej za generowanie par testowych ma podwojoną liczbę przerzutników. Do testowanej sieci n połączeń jest podłączony tylko co drugi przerzutnik. Taka struktura generuje wszystkie pary niezbędne do pobudzenia przesłuchów co jest niemożliwe w klasycznej strukturze R-LFSR. Eksperymenty potwierdziły skuteczność testera BIST w pobudzaniu określonych przesłuchów.A new method of detection of crosstalk faults is presented in the paper. An interconnect network employed by a target application is a sole subject of the test. The detection of crosstalk fault requires stimulation of the interconnect network under test (INUT) with two consecutive test patterns. The test patterns have to be applied to inputs of the INUT at a nominal clock frequency. So using the Built In Self Test (BIST) is a must. The proposed BIST structure is based on a ring register called 3n R LFSR (Fig.1). In contrast to a typical ring register, the 3n R LFSR contains a double number 2n of flip flops in its part that is responsible for two test pattern generation. The n lines of the INUT are fed from the outputs of every second flip flop of that part of the register. Such structure of the BIST is capable of generating all two test patterns that are required to stimulate crosstalk faults in the INUT, which is impossible in the case of a classical R LFSR. At the beginning of a test session the 3n-R-LFSR is seeded with a chosen value. After g clock cycles the final state (signature) is read. In more complex cases crosstalk can be observed only if a number k of lines being aggressors change their state simultaneously. The experiments proved that for k << n it is possible to find the initial seed being the beginning of a test sequence, that stimulate all required crosstalks. The length of the test sequence and simulation time ? necessary for finding initial seed is acceptable (Tab. 3)