Distinct regions of RPB11 are required for heterodimerization with RPB3 in human and yeast RNA polymerase II

In Saccharomyces cerevisiae, RNA polymerase II assembly is probably initiated by the formation of the RPB3–RPB11 heterodimer. RPB3 is encoded by a single copy gene in the yeast, mouse and human genomes. The RPB11 gene is also unique in yeast and mouse, but in humans a gene family has been identified that potentially encodes several RPB11 proteins differing mainly in their C-terminal regions. We compared the abilities of both yeast and human proteins to heterodimerize. We show that the yeast RPB3/RPB11 heterodimer critically depends on the presence of the C-terminal region of RPB11. In contrast, the human heterodimer tolerates significant changes in RPB11 C-terminus, allowing two human RPB11 variants to heterodimerize with the same efficiency with RPB3. In keeping with this observation, the interactions between the conserved N-terminal ‘α-motifs’ is much more important for heterodimerization of the human subunits than for those in yeast. These data indicate that the heterodimerization interfaces have been modified during the course of evolution to allow a recent diversification of the human RPB11 subunits that remains compatible with heterodimerization with RPB3

Ancient origin, functional conservation and fast evolution of DNA-dependent RNA polymerase III

RNA polymerase III contains seventeen subunits in yeasts (Saccharomyces cerevisiae and Schizosaccharomyces pombe) and in human cells. Twelve of them are akin to the core RNA polymerase I or II. The five other are RNA polymerase III-specific and form the functionally distinct groups Rpc31-Rpc34-Rpc82 and Rpc37-Rpc53. Currently sequenced eukaryotic genomes revealed significant homology to these seventeen subunits in Fungi, Animals, Plants and Amoebozoans. Except for subunit Rpc31, this also extended to the much more distantly related genomes of Alveolates and Excavates, indicating that the complex subunit organization of RNA polymerase III emerged at a very early stage of eukaryotic evolution. The Sch.pombe subunits were expressed in S.cerevisiae null mutants and tested for growth. Ten core subunits showed heterospecific complementation, but the two largest catalytic subunits (Rpc1 and Rpc2) and all five RNA polymerase III-specific subunits (Rpc82, Rpc53, Rpc37, Rpc34 and Rpc31) were non-functional. Three highly conserved RNA polymerase III-specific domains were found in the twelve-subunit core structure. They correspond to the Rpc17-Rpc25 dimer, involved in transcription initiation, to an N-terminal domain of the largest subunit Rpc1 important to anchor Rpc31, Rpc34 and Rpc82, and to a C-terminal domain of Rpc1 that presumably holds Rpc37, Rpc53 and their Rpc11 partner

A human RNA polymerase II subunit is encoded by a recently generated multigene family

BACKGROUND: The sequences encoding the yeast RNA polymerase II (RPB) subunits are single copy genes. RESULTS: While those characterized so far for the human (h) RPB are also unique, we show that hRPB subunit 11 (hRPB11) is encoded by a multigene family, mapping on chromosome 7 at loci p12, q11.23 and q22. We focused on two members of this family, hRPB11a and hRPB11b: the first encodes subunit hRPB11a, which represents the major RPB11 component of the mammalian RPB complex ; the second generates polypeptides hRPB11bα and hRPB11bβ through differential splicing of its transcript and shares homologies with components of the hPMS2L multigene family related to genes involved in mismatch-repair functions (MMR). Both hRPB11a and b genes are transcribed in all human tissues tested. Using an inter-species complementation assay, we show that only hRPB11bα is functional in yeast. In marked contrast, we found that the unique murine homolog of RPB11 gene maps on chromosome 5 (band G), and encodes a single polypeptide which is identical to subunit hRPB11a. CONCLUSIONS: The type hRPB11b gene appears to result from recent genomic recombination events in the evolution of primates, involving sequence elements related to the MMR apparatus

The archaeal RNA polymerase subunit P and the eukaryotic polymerase subunit Rpb12 are interchangeable in vivo and in vitro

The general subunit of all three eukaryotic RNA polymerases, Rpb12, and subunit P of the archaeal enzyme show sequence similarities in their N-terminal zinc ribbon and some highly conserved residues in the C-terminus. We report here that archaeal subunit P under the control of a strong yeast promoter could complement the lethal phenotype of a RPB12 deletion mutant and that subunit Rpb12 from yeast can functionally replace subunit P during reconstitution of the archaeal RNA polymerase. The ΔP enzyme is unable to form stable open complexes, but can efficiently extend a dinucleotide on a premelted template or RNA on an elongation scaffold. This suggests that subunit P is directly or indirectly involved in promoter opening. The activity of the ΔP enzyme can be rescued by the addition of Rpb12 or subunit P to transcription reactions. Mutation of cysteine residues in the zinc ribbon impair the activity of the enzyme in several assays and this mutated form of P is rapidly replaced by wild-type P in transcription reactions. The conserved zinc ribbon in the N-terminus seems to be important for proper interaction of the complete subunit with other RNA polymerase subunits and a 17-amino-acid C-terminal peptide is sufficient to support all basic RNA polymerase functions in vitro

РАСЧЕТ СТАТИЧЕСКИХ ПАРАМЕТРОВ КРЕМНИЕВОГО ДИОДА, СОДЕРЖАЩЕГО В СИММЕТРИЧНОМ p–n-ПЕРЕХОДЕ δ-СЛОЙ ТОЧЕЧНЫХ ТРЕХЗАРЯДНЫХ ДЕФЕКТОВ

The study of semiconductor materials and devices containing a narrow layer of impurity atoms and/or intrinsic point defects of the crystal lattice is of fundamental and practical interest. The aim of the study is to calculate the electric parameters of a symmetric silicon diode, in the flat p–n-junction of which a δ-layer of point triple-charged t-defects is formed. Such a diode is called p–t–n-diode, similarly to p–i–n-diode.Each t-defect can be in one of the three charge states (−1, 0, and +1; in the units of the elementary charge). It is assumed that at room temperature all hydrogen-like acceptors in p-region and hydrogen-like donors in n-region are ionized. It was assumed that the cross-section for v-band hole capture on t-defects is greater than the cross-section for c-band electron capture on t-defects.The system of stationary nonlinear differential equations, which describe in the drift-diffusion approximation a migration of electrons and holes in semiconductors, is solved numerically. The static capacityvoltage and current-voltage characteristics of the silicon diode with nondegenerate regions of pand n-type of electrical conductivity are calculated for forward and reverse electric bias voltage.It is shown by calculation that in the p–t–n-diode containing the δ-layer of t-defects, at the forward bias a region of current density stabilization occurs. At the reverse bias the current density in such a diode is much greater than the one in a p–n-diode without t-defects. With the reverse bias the capacitance of the p–t–n-diode, in contrast to the p–n-diode, increases at first and then decreases.Научный и практический интерес представляет изучение полупроводниковых материалов и приборов с узким слоем атомов примесей и/или собственных точечных дефектов кристаллической решетки. Цель работы – рассчитать электрические параметры симметричного кремниевого диода, в плоском p–n-переходе которого сформирован δ-слой точечных трехзарядных t-дефектов. Такой диод называется p–t–n-диодом, подобно p–i–n-диоду.Каждый t-дефект может находиться в одном из трех зарядовых состояний (−1, 0, +1; в единицах элементарного заряда). Считается, что при комнатной температуре все водородоподобные акцепторы в p-области и водородоподобные доноры в n-области ионизованы. Принималось, что сечение захвата дырок v-зоны на t-дефекты больше сечения захвата электронов c-зоны на t-дефекты.Численно решена система cтационарных нелинейных дифференциальных уравнений, описывающих в дрейфово-диффузионном приближении миграцию электронов и дырок в полупроводниках. Рассчитаны статические вольт-фарадные и вольт-амперные характеристики кремниевого диода с невырожденными областями p- и n-типа электропроводности при прямом и обратном электрическом напряжении смещения.Расчетным путем показано, что в p–t–n-диоде, содержащем δ-слой t-дефектов, при прямом смещении имеется участок стабилизации плотности тока. При обратном смещении плотность тока в таком диоде много больше, чем в p–n-диоде без t-дефектов. При увеличении обратного смещения емкость p–t–n-диода, в отличие от p–n-диода, вначале увеличивается, а затем уменьшается

Контроль дифференциального сопротивления p–n-переходов биполярного транзистора в активном режиме методом импедансной спектроскопии

Controlling of parameters of manufactured transistors and interoperational controlling during their production are necessary conditions for production of competitive products of electronic industry. Traditionally for controlling of bipolar transistors the direct current measurements and registration of capacity-voltage characteristics are used. Carrying out measurements on alternating current in a wide interval of frequencies (20 Hz–30 MHz) will allow to obtain additional information on parameters of bipolar transistors. The purpose of the work is to show the possibilities of the method of impedance spectroscopy for controlling of differential resistance of p–n-junctions of the bipolar p–n–p-transistor in active mode.The KT814G p–n–p-transistor manufactured by JSC “INTEGRAL” was studied by the method of impedance spectroscopy. The values of differential electrical resistance and capacitance for base–emitter and base–collector p–n-junctions are defi at direct currents in base from 0.8 to 46 µA.The results of the work can be applied to elaboration of techniques of fi checking of discrete bipolar semiconductor devices.Контроль параметров готовых транзисторов и межоперационный контроль при их изготовлении являются необходимыми условия выпуска конкурентоспособных изделий электронной промышленности. Традиционно для контроля биполярных транзисторов используются измерения на постоянном токе и регистрация вольт-фарадных характеристик. Проведение измерений на переменном токе позволит получить дополнительную информацию о параметрах биполярных транзисторов.Цель работы – показать возможности метода импедансной спектроскопии для контроля дифференциального электрического сопротивления p–n-переходов биполярного p–n–p-транзистора в активном режиме.Методом импедансной спектроскопии исследован p–n–p-транзистор КТ814Г производства ОАО «ИНТЕГРАЛ». На переменном токе в интервале частот 20 Hz–30 MHz определены значения дифференциального электрического сопротивления и емкости p–n-переходов база–эмиттера и база–коллектора при постоянных токах базы от 0,8 до 46 µA.Результаты работы могут быть использованы при отработке методик выходного контроля дискретных биполярных полупроводниковых приборов

Влияние экстракции дырок из базовой области кремниевого p–n–p-транзистора на его реактивный импеданс

Transistor structures are the basic elements of integrated circuitry and are often used to create not only transistors themselves, but also diodes, resistors, and capacitors. Determining the mechanism of the occurrence of inductive type impedance in semiconductor structures is an urgent task, the solution of which will create the prerequisites for the development of solid-state analogs of inductors. The purpose of the work is to establish the effect of extraction of non-equilibrium charge carriers from the base region on the reactive impedance of a bipolar p–n–p transistor.Using impedance spectroscopy in the frequency range 20 Hz–30 MHz, the structures based on p–n–p transistors KT814G manufactured by JSC “INTEGRAL” were studied. It is shown that in the transistor structures it is possible to observe the “effect of negative capacitance” (inductive type impedance). It is established that the most probable cause of the inductive type impedance is the accumulation of uncompensated charge of holes in the base, the value of inductive impedance is influenced by both the injection efficiency in the base–emitter junction and the extraction efficiency in the base–collector junction.The results can be applied in the elaboration of technologies for the formation of elements of silicon based integrated circuits with an impedance of inductive type.Транзисторные структуры являются базовыми элементами интегральной схемотехники и часто используются для создания не только собственно транзисторов, но и диодов, резисторов, конденсаторов. Определение механизма возникновения импеданса индуктивного типа в полупроводниковых структурах является актуальной задачей, решение которой создаст предпосылки к разработке твердотельных аналогов катушек индуктивности. Цель работы – установить влияние экстракции неравновесных носителей заряда из базовой области на реактивный импеданс биполярного p–n–p-транзистора.Методом импедансной спектроскопии в интервале частот 20 Hz–30 MHz исследованы структуры на базе p–n–p-транзисторов КТ814Г производства ОАО «ИНТЕГРАЛ». Показано, что в транзисторных структурах возможно наблюдение «эффекта отрицательной ёмкости» (импеданс индуктивного типа). Установлено, что наиболее вероятной причиной возникновения импеданса индуктивного типа является накопление нескомпенсированного заряда дырок в базе, а на величину индуктивного импеданса влияет как эффективность инжекции в переходе база–эмиттер, так и эффективность экстракции в переходе база–коллектор.Результаты работы могут быть использованы при разработке технологий формирования элементов интегральных микросхем на основе кремния с импедансом индуктивного типа

The fission yeast Rpb4 subunit of RNA polymerase II plays a specialized role in cell separation

RNA polymerase II is a complex of 12 subunits, Rpb1 to Rpb12, whose specific roles are only partly understood. Rpb4 is essential in mammals and fission yeast, but not in budding yeast. To learn more about the roles of Rpb4, we expressed the rpb4 gene under the control of regulatable promoters of different strength in fission yeast. We demonstrate that below a critical level of transcription, Rpb4 affects cellular growth proportional to its expression levels: cells expressing lower levels of rpb4 grew slower compared to cells expressing higher levels. Lowered rpb4 expression did not affect cell survival under several stress conditions, but it caused specific defects in cell separation similar to sep mutants. Microarray analysis revealed that lowered rpb4 expression causes a global reduction in gene expression, but the transcript levels of a distinct subset of genes were particularly responsive to changes in rpb4 expression. These genes show some overlap with those regulated by the Sep1-Ace2 transcriptional cascade required for cell separation. Most notably, the gene expression signature of cells with lowered rpb4 expression was highly similar to those of mcs6, pmh1, sep10 and sep15 mutants. Mcs6 and Pmh1 encode orthologs of metazoan TFIIH-associated cyclin-dependent kinase (CDK)-activating kinase (Cdk7-cyclin H-Mat1), while Sep10 and Sep15 encode mediator components. Our results suggest that Rpb4, along with some other general transcription factors, plays a specialized role in a transcriptional pathway that controls the cell cycle-regulated transcription of a specific subset of genes involved in cell division. ELECTRONIC SUPPLEMENTARY MATERIAL: Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00438-006-0161-5 and is accessible for authorized users

СПЕКТРЫ DLTS КРЕМНИЕВЫХ ДИОДОВ С p+—n–ПЕРЕХОДОМ, ОБЛУЧЕННЫХ ВЫСОКОЭНЕРГЕТИЧЕСКИМИ ИОНАМИ КРИПТОНА

p+-n-Diodes have been studied. The diodes were manufactured on wafers (thickness 460 μm, (111) plane) of uniformly phosphorus doped float–zone–grown single–crystal silicon. The resistivity of silicon was 90 Ohm · cm and the phosphorus concentration was 5 · 1013 cm–3. The diodes were irradiated with 250 MeV krypton ions. The irradiation fluence was 108 cm–2. Deep–level transient spectroscopy (DLTS) was used to examine the defects induced by high energy krypton ion implantation. The DLTS spectra were recorded at a frequency of 1 MHz in the 78—290 K temperature range. The capacity–voltage characteristics have been measured at a reverse bias voltage from 0 to –19 V at a frequency of 1 MHz. We show that the main irradiation–induced defects are A–centers and divacancies. The behavior of DLTS spectra in the 150—260 K temperature range depends essentially on the emission voltage Ue. The variation of Ue allows us to separate the contributions of different defects into the DLTS spectrum in the 150—260 K temperature range. We show that, in addition to A–centers and divacancies, irradiation produces multivacancy complexes with the energy level Et = Ec – (0.5 ± 0.02) eV and an electron capture cross section of ~4 · 10–13 cm2.Исследованы p+—n-диоды. Диоды изготовлены на пластинах однородно легированного фосфором монокристаллического кремния (толщина 460 мкм, плоскость (111)), выращенного методом бестигельной зонной плавки. Удельное сопротивление кремния — 90 Ом × см, концентрация фосфора — 5 × 1013 см−3. Диоды подвергнуты облучению ионами криптона с энергией 250 МэВ. Флюенс облучения — 108 см−2. Радиационные дефекты, вводимые высокоэнергетической имплантацией ионов криптона, исследованы с помощью нестационарной спектроскопии глубоких уровней (DLTS — Deep−level transient spectroscopy). Спектры DLTS регистрировали на частоте 1 МГц в интервале температур 78—290 К. Вольт-фарадные характеристики измерены при напряжении обратного смещения от 0 до – 19 В на частоте 1 МГц. Показано, что основными радиационными дефектами являются А−центры и дивакансии. Установлено, что вид спектров DLTS в интервале температур 150—260 K существенно зависит от напряжения эмиссии Ue. Варьирование Ue в ходе эксперимента позволило разделить вклады от различных дефектов в спектр DLTS в интервале температур 150—260 К. Показано, что, помимо А−центров и дивакансий, при облучении формируются многовакансионные комплексы с энергетическим уровнем Et = Ec -(0,50 ± 0,02) эВ и сечением захвата электронов ~ 4 × 10−13 см2