Students Helping Students

A Montana Tech student was injured and his home needed to be made wheelchair accessible. Highlands College Construction Technology students designed and constructed an exterior ramp leading to the house as well as interior ramps between levels inside the house.

INVESTIGATION OF STRATEGIES TO PROMOTE EFFECTIVE TEACHER PROFESSIONAL DEVELOPMENT EXPERIENCES IN EARTH SCIENCE

This dissertation serves as a call to geoscientists to share responsibility with K-12 educators for increasing Earth science literacy. When partnerships are created among K-12 educators and geoscientists, the synergy created can promote Earth science literacy in students, teachers, and the broader community. The research described here resulted in development of tools that can support effective professional development for teachers. One tool is used during the planning stages to structure a professional development program, another set of tools supports measurement of the effectiveness of a development program, and the third tool supports sustainability of professional development programs. The Michigan Teacher Excellence Program (MiTEP), a Math/Science Partnership project funded by the National Science Foundation, served as the test bed for developing and testing these tools. The first tool, the planning tool, is the Earth Science Literacy Principles (ESLP). The ESLP served as a planning tool for the two-week summer field courses as part of the MiTEP program. The ESLP, published in 2009, clearly describe what an Earth science literate person should know. The ESLP consists of nine big ideas and their supporting fundamental concepts. Using the ESLP for planning a professional development program assisted both instructors and teacher-participants focus on important concepts throughout the professional development activity. The measurement tools were developed to measure change in teachers’ Earth science content-area knowledge and perceptions related to teaching and learning that result from participating in a professional development program. The first measurement tool, the Earth System Concept Inventory (ESCI), directly measures content-area knowledge through a succession of multiple-choice questions that are aligned with the content of the professional development experience. The second measurement, an exit survey, collects qualitative data from teachers regarding their impression of the professional development. Both the ESCI and the exit survey were tested for validity and reliability. Lesson study is discussed here as a strategy for sustaining professional development in a school or a district after the end of a professional development activity. Lesson study, as described here, was offered as a formal course. Teachers engaged in lesson study worked collaboratively to design and test lessons that improve the teachers’ classroom practices. Data regarding the impact of the lesson study activity were acquired through surveys, written documents, and group interviews. The data are interpreted to indicate that the lesson study process improved teacher quality and classroom practices. In the case described here, the lesson study process was adopted by the teachers’ district and currently serves as part of the district’s work in Professional Learning Communities, resulting in ongoing professional development throughout the district.

Diastolic dysfunction of the cardiac allograft and maximal exercise capacity

BACKGROUND: Peak oxygen uptake (peak Vo(2)) is an established integrative measurement of maximal exercise capacity in cardiovascular disease. After heart transplantation (HTx) peak Vo(2) remains reduced despite normal systolic left ventricular function, which highlights the relevance of diastolic function. In this study we aim to characterize the predictive significance of cardiac allograft diastolic function for peak Vo(2). METHODS: Peak Vo(2) was measured using a ramp protocol on a bicycle ergometer. Left ventricular (LV) diastolic function was assessed with tissue Doppler imaging sizing the velocity of the early (Ea) and late (Aa) apical movement of the mitral annulus, and conventional Doppler measuring early (E) and late (A) diastolic transmitral flow propagation. Correlation coefficients were calculated and linear regression models fitted. RESULTS: The post-transplant time interval of the 39 HTxs ranged from 0.4 to 20.1 years. The mean age of the recipients was 55 +/- 14 years and body mass index (BMI) was 25.4 +/- 3.9 kg/m(2). Mean LV ejection fraction was 62 +/- 4%, mean LV mass index 108 +/- 22 g/m(2) and mean peak Vo(2) 20.1 +/- 6.3 ml/kg/min. Peak Vo(2) was reduced in patients with more severe diastolic dysfunction (pseudonormal or restrictive transmitral inflow pattern), or when E/Ea was > or =10. Peak Vo(2) correlated with recipient age (r = -0.643, p < 0.001), peak heart rate (r = 0.616, p < 0.001) and BMI (r = -0.417, p = 0.008). Of all echocardiographic measurements, Ea (r = 0.561, p < 0.001) and Ea/Aa (r = 0.495, p = 0.002) correlated best. Multivariate analysis identified age, heart rate, BMI and Ea/Aa as independent predictors of peak Vo(2). CONCLUSIONS: Diastolic dysfunction is relevant for the limitation of maximal exercise capacity after HTx.

Seasonal variation of VO 2 max and the VO2-work rate relationship in elite Alpine skiers

PURPOSE: Alpine ski performance relates closely to both anaerobic and aerobic capacities. During their competitive season, skiers greatly reduce endurance and weight training, and on-snow training becomes predominant. To typify this shift, we compared exhaustive ramp cycling and squat (SJ) and countermovement jumping (CMJ) performance in elite males before and after their competitive season. RESULTS: In postseason compared with preseason: 1) maximal oxygen uptake (VO 2 max) normalized to bodyweight was higher (55.2 +/- 5.2 vs 52.7 +/- 3.6 mL x kg(-1) x min(-1), P < 0.01), but corresponding work rate (W) was unchanged; 2) at ventilatory thresholds (VT), absolute and relative work rates were similar but heart rates were lower; 3) VO2/W slope was greater (9.59 +/- 0.6 vs 9.19 +/- 0.4 mL O2 x min(-1) x W(-1), P = 0.02), with similar flattening (P < 0.01) above V T1 at both time points; and 4) jump height was greater in SJ (47.4 +/- 4.4 vs 44.7 +/- 4.3 cm, P < 0.01) and CMJ (52.7 +/- 4.6 vs 50.4 +/- 5.0 cm, P < 0.01). DISCUSSION: We believe that aerobic capacity and leg power were constrained in preseason and that improvements primarily reflected an in-season recovery from a fatigued state, which was caused by incongruous preseason training. Residual adaptations to high-altitude exposure in preseason could have also affected the results. Nonetheless, modern alpine skiing seemingly provides an ample cardiovascular training stimulus for skiers to maintain their aerobic capacities during the racing season. CONCLUSIONS: We conclude that aerobic fitness and leg explosiveness can be maintained in-season but may be compromised by heavy or excessive preseason training. In addition, ramp test V O2/W slope analysis could be useful for monitoring both positive and negative responses to training.

Modifying peripheral IV catheters with side holes and side slits results in favorable changes in fluid dynamic properties during the injection of iodinated contrast material

OBJECTIVE: The purpose of this study was to compare a standard peripheral end-hole angiocatheter with those modified with side holes or side slits using experimental optical techniques to qualitatively compare the contrast material exit jets and using numeric techniques to provide flow visualization and quantitative comparisons. MATERIALS AND METHODS: A Schlieren imaging system was used to visualize the angiocatheter exit jet fluid dynamics at two different flow rates. Catheters were modified by drilling through-and-through side holes or by cutting slits into the catheters. A commercial computational fluid dynamics package was used to calculate numeric results for various vessel diameters and catheter orientations. RESULTS: Experimental images showed that modifying standard peripheral IV angiocatheters with side holes or side slits qualitatively changed the overall flow field and caused the exiting jet to become less well defined. Numeric calculations showed that the addition of side holes or slits resulted in a 9-30% reduction of the velocity of contrast material exiting the end hole of the angiocatheter. With the catheter tip directed obliquely to the wall, the maximum wall shear stress was always highest for the unmodified catheter and was always lowest for the four-side-slit catheter. CONCLUSION: Modified angiocatheters may have the potential to reduce extravasation events in patients by reducing vessel wall shear stress.

Physiological response to increasing levels of neurally adjusted ventilatory assist (NAVA)

This study evaluated the response to increasing levels of neurally adjusted ventilatory assist (NAVA), a mode converting electrical activity of the diaphragm (EAdi) into pressure, regulated by a proportionality constant called the NAVA level. Fourteen rabbits were studied during baseline, resistive loading and ramp increases of the NAVA level. EAdi, airway (Paw) and esophageal pressure (Pes), Pes pressure time product (PTPes), breathing pattern, and blood gases were measured. Resistive loading increased PTPes and EAdi. P(a)(CO)(2) increased with high load but not during low load. Increasing NAVA levels increased Paw until a breakpoint where the Paw increase was reduced despite increasing NAVA level. At this breakpoint, Pes, PTPes, EAdi, and P(a)(CO)(2) were similar to baseline. Further increase of the NAVA level reduced Pes, PTPes and EAdi without changes in ventilation. In conclusion, observing the trend in Paw during a ramp increase of the NAVA level allows determination of a level where the inspiratory effort matches unloaded conditions.

Compound heterozygous mutations affect protein folding and function in patients with congenital sucrase-isomaltase deficiency

BACKGROUND & AIMS: Congenital sucrase-isomaltase (SI) deficiency is an autosomal-recessive intestinal disorder characterized by a drastic reduction or absence of sucrase and isomaltase activities. Previous studies have indicated that single mutations underlie individual phenotypes of the disease. We investigated whether compound heterozygous mutations, observed in some patients, have a role in disease pathogenesis. METHODS: We introduced mutations into the SI complementary DNA that resulted in the amino acid substitutions V577G and G1073D (heterozygous mutations found in one group of patients) or C1229Y and F1745C (heterozygous mutations found in another group). The mutant genes were expressed transiently, alone or in combination, in COS cells and the effects were assessed at the protein, structural, and subcellular levels. RESULTS: The mutants SI-V577G, SI-G1073D, and SI-F1745C were misfolded and could not exit the endoplasmic reticulum, whereas SI-C1229Y was transported only to the Golgi apparatus. Co-expression of mutants found on each SI allele in patients did not alter the protein's biosynthetic features or improve its enzymatic activity. Importantly, the mutations C1229Y and F1745C, which lie in the sucrase domains of SI, prevented its targeting to the cell's apical membrane but did not affect protein folding or isomaltase activity. CONCLUSIONS: Compound heterozygosity is a novel pathogenic mechanism of congenital SI deficiency. The effects of mutations in the sucrase domain of SIC1229Y and SIF1745C indicate the importance of a direct interaction between isomaltase and sucrose and the role of sucrose as an intermolecular chaperone in the intracellular transport of SI.

Traffic Routes and Signals for the Tonoplast

Tonoplast, the membrane delimiting plant vacuoles, regulates ion, water and nutrient movement between the cytosol and the vacuolar lumen through the activity of its membrane proteins. Correct traffic of proteins from the endoplasmic reticulum (ER) to the tonoplast requires (i) approval by the ER quality control, (ii) motifs for exit from the ER and (iii) motifs that promote sorting to the tonoplast. Recent evidence suggests that this traffic follows different pathways that are protein-specific and could also reflect vacuole specialization for lytic or storage function. The routes can be distinguished based on their sensitivity to drugs such as brefeldin A and C834 as well as using mutant plants that are defective in adaptor proteins of vesicle coats, or dominant-negative mutants of Rab GTPases.

Sportvereine als Interessenorganisationen?! Ziel-Interessen-Divergenzen und ihre Auswirkungen

Einleitung: Freiwillige Sportorganisationen wie Sportvereine sind als Interessenorganisationen zu charakterisieren, deren primärer Organisationszweck sich auf die Vertretung der Interessen ihrer Mitglieder bezieht (Nagel, 2006). Mit dieser Interessenvertretungsfunktion wird gewährleistet, dass die Vereinsziele an die Mitgliederinteressen rückgebunden bleiben und Ziel-Interessen-Kongruenz herrscht (Schimank, 2005). Die Beobachtungen der Ver-einsrealität weisen jedoch darauf hin, dass die Ziele der Vereine oftmals von den Interessen ihrer Mitglieder abweichen und im Widerspruch zueinander stehen (Baur & Burrmann, 2003; Heinemann, 2004). Vor diesem Problemhintergrund ist zu fragen: Inwiefern stimmen die Vereinsziele mit den Mitgliederinteressen überein? Und für den Fall von Ziel-Interessen-Divergenzen, welche Auswirkungen ergeben sich daraus für die Vereine? Theoretisch-methodischer Ansatz: Für die theoriegeleite Beantwortung der Fragestellungen wurde auf Grundlage einer akteurtheoretischen Kon-zeptualisierung von Vereinen als Interessenorganisationen (Nagel, 2006; Schimank, 2007) ein strukturell-individualistisches Mehr-Ebenen-Modell entwickelt, dass zum einen das Verhältnis von Vereinszielen (Vereins-ebene) und Mitgliederinteressen (Mitgliederebene) und damit die Genese von Ziel-Interessen-Divergenzen erklärt. Zum anderen liefert das Modell eine Erklärungsgrundlage, welche Auswirkungen Divergenzen (struktu-relle Vereinssituation) auf die Mitglieder haben, d.h. wie die Mitglieder auf bestehende Divergenzen reagieren (individuelle Mitgliederhandlung). Die erforderlichen Vereins- und Mitgliederdaten wurden im Rahmen zweier Teilstudien erfasst, bei denen in erster Linie quantitative Erhebungs- und Auswertungsverfahren zum Einsatz kamen. Die erste Teilstudie wurde im Fürstentum Liechtenstein zur Analyse der Ziel-Interessen-Übereinstimmung durchgeführt, wobei hierzu die Präsidenten von 17 Vereinen und 481 Mitglieder schriftlich befragt wurden. In der zweiten Teilstudie wurde im Schweizer Turnverein Länggasse Bern mit 199 Mitgliedern eine schriftliche Befragung durchgeführt, um die Auswirkungen von Divergenzen zu analysieren. Zur Erfassung des Ausmasses von Ziel-Interessen-Divergenzen wurde eine Vergleichsprüfung der Vereinsziele und Mitgliederinteressen durchgeführt. Die Liechtensteinischen Vereinspräsidenten und Vereinsmitglieder mussten diesbezüglich die Bedeutung von 23 verschiedenen Items eines entwickelten Ziel-/Interessen-Fragekataloges einschätzen und über den betragsmässigen Bedeutungsunterschied konnte das quantitative Divergenzmass bestimmt werden. Um die Frage nach den Auswirkungen zu klären, hatten die Mitglieder des Schweizer Turnvereins Länggase Bern Aussagen über ihre Reaktionen auf bestehende Divergenzen entlang einer konzipierten Handlungstypologie (exit, voice, loyalty, neglect) zu geben. Ergebnisse: Die Befunde der Vergleichsanalyse zeigen, dass in allen untersuchten Vereinen Ziel-Interessen-Divergenzen in unterschiedlichem Ausmaß existieren. Bei einem Drittel der Vereine bestehen mittlere Divergenzen und in zwei Drittel sind größere Divergenzen zu beobachten. Divergenzen bestehen dabei vor allem hinsichtlich des Brei-tensports (v.a. Erweiterung des Sportangebots), der Vereinsentwicklung (v.a. Öffnen für neue Entwicklungen) und der Geselligkeit (v.a. aussersportliches Angebot, Wahrung der Vereinstradition). Die Analyse der Auswirkungen zeigt, dass geringe und mittelstarke Divergenzen in erster Linie zu konstruktiven Reaktionen führen. Die Mitglieder machen dabei die Vereinsführung auf diese Situation aufmerksam (voice), indem sie das Problem ansprechen und Vorschläge zur Problemlösung unterbreiten. Oder sie haben aus Grün-den der Vereinsverbundenheit (loyalty) die Hoffnung und das Vertrauen, dass die Vereinsführung ohne ihr Eingreifen das Problem löst. Wenn aber großen Divergenzen existieren, die sich für die Vereine zu einem gra-vierenden Problem entwickeln, tendieren die Mitglieder hingegen verstärkt zu destruktiven Reaktionen, die sich durch Gleichgültigkeit und Desinteresse am Vereinsgeschehen (neglect) äussern, indem die Mitglieder zukünftig nicht mehr auf Vereinsanlässe gehen. Oder sie wählen die Option des Rückzugs und Austritts (exit), indem sie ihr Vereinsengagement aufgeben oder im Extremfall aus der Abteilung oder dem Verein austreten. Diskussion: In der Vereinsrealität stellen Ziel-Interessen-Divergenzen offenbar eher die Norm als die Ausnahme dar, so dass die Vereinsziele nur teilweise an die Mitgliederinteressen rückgebunden sind. Allerdings sind Divergenzen nicht grundsätzlich als Problem zu interpretieren, sondern sind vielmehr in Abhängigkeit ihrer Intensität zu beurtei-len. Diesbezüglich dürften Vereine geringfügig bis mittelmässig ausgeprägte Divergenzsituationen im laufenden Vereinsgeschehen bis zu einem gewissen Grad relativ gut aushalten und aufarbeiten können, weil die Mitglie-der in diesem Fall zu einem konstruktiven Umgang tendieren, die eine Wiederherstellung von Ziel-Interessen-Kongruenz begünstigt. Demgegenüber können sich aber grosse Divergenzen zu einem ernsthaften Problem für Vereine entfalten, weil die Mitglieder in diesem Fall zu destruktiven Reaktionen neigen, die sich negativ auf die Vereinsentwicklung auswirken dürfte. Literatur Baur, J. & Burrmann, U. (2003). Konflikte in Sportvereinen. In J. Baur & S. Braun (Hrsg.), Integrationsleistungen von Sport-vereinen als Freiwilligenorganisationen (S. 331-370). Aachen: Meyer & Meyer. Nagel, S. (2006). Sportvereine im Wandel. Akteurtheoretische Analysen zur Entwicklung von Sportvereinen. Schorndorf: Hofmann. Nagel, S. (2007). Akteurtheoretische Analyse der Sportvereinsentwicklung. Ein theoretisch-methodischer Bezugsrahmen. Sportwissenschaft, 37 (2), 186-201. Schimank, U. (2005). Der Vereinssport in der Organisationsgesellschaft: organisationssoziologische Perspektiven auf ein spannungsreiches Verhältnis. In T. Alkemeyer, B. Rigauer & G. Sobiech (Hrsg.), Organisationsentwicklungen und De-Institutionalisierungsprozesse im Sport (S. 21-44). Schorndorf: Hofmann. Schimank, U. (2007). Handeln und Strukturen. Einführung in die akteurtheoretische Soziologie. Weinheim: Juventa Verlag.

Mammalian iron transporters: Families SLC11 and SLC40

This review is focused on the mammalian SLC11 and SLC40 families and their roles in iron homeostasis. The SLC11 family is composed of two members, SLC11A1 and SLC11A2. SLC11A1 is expressed in the lysosomal compartment of macrophages and in the tertiary granules of neutrophils, playing a key role in innate resistance against infection by intracellular microbes. SLC11A2 is a key player in iron metabolism and is ubiquitously expressed, most notably in the proximal duodenum, immature erythroid cells, brain, placenta and kidney. Intestinal iron absorption is mediated by SLC11A2 at the apical membrane of enterocytes, followed by basolateral exit via SLC40A1. To meet the daily requirement for iron, approximately 80% of the iron comes from the breakdown of hemoglobin following macrophage phagocytosis of senescent erythrocytes (iron recycling). Both SLC11A1 and SLC11A2 play an important role in macrophage iron recycling. SLC11A2 also transports iron into the cytosol across the membrane of endocytotic vesicles of the transferrin receptor-cycle. SLC40A1 is the sole member of the SLC40 family and is involved in the only cellular iron efflux mechanism described. SLC40A1 is highly expressed in several tissues and cells that play a critical role in body iron homeostasis. The signaling pathways that regulate SLC11A2 and SLC40A1 expression at transcriptional, post-transcriptional and post-translational levels are discussed. The roles of SLC11A2 and/or SLC40A1 in iron-associated disorders such as hemochromatosis, neurodegenerative diseases, and breast cancer are also summarized.

THE SOLAR WIND AS A POSSIBLE SOURCE OF FAST TEMPORAL VARIATIONS OF THE HELIOSPHERIC RIBBON

We present a possible source of pickup ions (PUIs) the ribbon observed by the Interstellar Boundary EXplorer (IBEX). We suggest that a gyrating solar wind and PUIs in the ramp and in the near downstream region of the termination shock (TS) could provide a significant source of energetic neutral atoms (ENAs) in the ribbon. A fraction of the solar wind and PUIs are reflected and energized during the first contact with the TS. Some of the solar wind may be reflected propagating toward the Sun but most of the solar wind ions form a gyrating beam-like distribution that persists until it is fully thermalized further downstream. Depending on the strength of the shock, these gyrating distributions can exist for many gyration periods until they are scattered/thermalized due to wave-particle interactions at the TS and downstream in the heliosheath. During this time, ENAs can be produced by charge exchange of interstellar neutral atoms with the gyrating ions. In order to determine the flux of energetic ions, we estimate the solar wind flux at the TS using pressure estimates inferred from in situ measurements. Assuming an average path length in the radial direction of the order of a few AU before the distribution of gyrating ions is thermalized, one can explain a significant fraction of the intensity of ENAs in the ribbon observed by IBEX. With a localized source and such a short integration path, this model would also allow fast time variations of the ENA flux.

NEW TARGET GENES FOR TUMOR SUPPRESSORS p53 AND p73 IN REGENERATING LIVER

The p53-family of proteins regulates expression of target genes during tissue development and differentiation. Within the p53-family, p53 and p73 have hepatic-specific functions in development and tumor suppression. Despite a growing list of p53/p73 target genes, very few of these have been studied in vivo, and the knowledge regarding functions of p53 and p73 in normal tissues remains limited. p53+/-p73+/- mice develop hepatocellular carcinoma (HCC), whereas overexpression of p53 in human HCC leads to tumor regression. However, the mechanism of p53/p73 function in liver remains poorly characterized. Here, the model of mouse liver regeneration is used to identify new target genes for p53/p73 in normal quiescent vs. proliferating cells. In response to surgical removal of ~2/3 of liver mass (partial hepatectomy, PH), the remaining hepatocytes exit G0 of cell cycle and undergo proliferation to reestablish liver mass. The hypothesis tested in this work is that p53/p73 functions in cell cycle arrest, apoptosis and senescence are repressed during liver regeneration, and reactivated at the end of the regenerative response. Chromatin immunoprecipitation (ChIP), with a p73-antibody, was used to probe arrayed genomic sequences (ChIP-chip) and uncover 158 potential targets of p73-regulation in normal liver. Global microarray analysis of mRNA levels, at T=0-48h following PH, revealed sets of genes that change expression during regeneration. Eighteen p73-bound genes changed expression after PH. Four of these genes, Foxo3, Jak1, Pea15, and Tuba1 have p53 response elements (p53REs), identified in silico within the upstream regulatory region. Forkhead transcription factor Foxo3 is the most responsive gene among transcription factors with altered expression during regenerative, cellular proliferation. p53 and p73 bind a Foxo3 p53RE and maintain active expression in quiescent liver. During liver regeneration, binding of p53 and p73, recruitment of acetyltransferase p300, and an active chromatin structure of Foxo3 are disrupted, alongside loss of Foxo3 expression. These parameters of Foxo3 regulation are reestablished at completion of liver growth and regeneration, supporting a temporary suspension of p53 and p73 regulatory functions in normal cells during tissue regeneration.

Structure of the Hsp110:Hsc70 nucleotide exchange machine.

Hsp70s mediate protein folding, translocation, and macromolecular complex remodeling reactions. Their activities are regulated by proteins that exchange ADP for ATP from the nucleotide-binding domain (NBD) of the Hsp70. These nucleotide exchange factors (NEFs) include the Hsp110s, which are themselves members of the Hsp70 family. We report the structure of an Hsp110:Hsc70 nucleotide exchange complex. The complex is characterized by extensive protein:protein interactions and symmetric bridging interactions between the nucleotides bound in each partner protein's NBD. An electropositive pore allows nucleotides to enter and exit the complex. The role of nucleotides in complex formation and dissociation, and the effects of the protein:protein interactions on nucleotide exchange, can be understood in terms of the coupled effects of the nucleotides and protein:protein interactions on the open-closed isomerization of the NBDs. The symmetrical interactions in the complex may model other Hsp70 family heterodimers in which two Hsp70s reciprocally act as NEFs.

Deadenylation is prerequisite for P-body formation and mRNA decay in mammalian cells.

Deadenylation is the major step triggering mammalian mRNA decay. One consequence of deadenylation is the formation of nontranslatable messenger RNA (mRNA) protein complexes (messenger ribonucleoproteins [mRNPs]). Nontranslatable mRNPs may accumulate in P-bodies, which contain factors involved in translation repression, decapping, and 5'-to-3' degradation. We demonstrate that deadenylation is required for mammalian P-body formation and mRNA decay. We identify Pan2, Pan3, and Caf1 deadenylases as new P-body components and show that Pan3 helps recruit Pan2, Ccr4, and Caf1 to P-bodies. Pan3 knockdown causes a reduction of P-bodies and has differential effects on mRNA decay. Knocking down Caf1 or overexpressing a Caf1 catalytically inactive mutant impairs deadenylation and mRNA decay. P-bodies are not detected when deadenylation is blocked and are restored when the blockage is released. When deadenylation is impaired, P-body formation is not restorable, even when mRNAs exit the translating pool. These results support a dynamic interplay among deadenylation, mRNP remodeling, and P-body formation in selective decay of mammalian mRNA.

Deadenylation is prerequisite for P-body formation and mRNA decay in mammalian cells.

Deadenylation is the major step triggering mammalian mRNA decay. One consequence of deadenylation is the formation of nontranslatable messenger RNA (mRNA) protein complexes (messenger ribonucleoproteins [mRNPs]). Nontranslatable mRNPs may accumulate in P-bodies, which contain factors involved in translation repression, decapping, and 5'-to-3' degradation. We demonstrate that deadenylation is required for mammalian P-body formation and mRNA decay. We identify Pan2, Pan3, and Caf1 deadenylases as new P-body components and show that Pan3 helps recruit Pan2, Ccr4, and Caf1 to P-bodies. Pan3 knockdown causes a reduction of P-bodies and has differential effects on mRNA decay. Knocking down Caf1 or overexpressing a Caf1 catalytically inactive mutant impairs deadenylation and mRNA decay. P-bodies are not detected when deadenylation is blocked and are restored when the blockage is released. When deadenylation is impaired, P-body formation is not restorable, even when mRNAs exit the translating pool. These results support a dynamic interplay among deadenylation, mRNP remodeling, and P-body formation in selective decay of mammalian mRNA.