A 42 kb region on human chromosome 9p21 encodes for three distinct tumor suppressors, p16INK4A, p14ARF and p15INK4B, and is altered in an estimated 30–40% of human tumors. The expression of the INK4A-ARF-INK4B gene cluster is silenced by polycomb during normal cell growth and is activated by oncogenic insults and during aging. How the polycomb is recruited to repress this gene cluster is unclear. Here, we show that expression of oncogenic Ras, which stimulates the expression of p15INK4B and p16INK4A, but not p14ARF, inhibits the expression of ANRIL (antisense non-coding RNA in the INK4 locus), a 3.8 kb-long non-coding RNA expressed in the opposite direction from INK4A-ARF-INK4B. We show that the p15INK4B locus is bound by SUZ12, a component of polycomb repression complex 2 (PRC2), and is H3K27-trimethylated. Notably, depletion of ANRIL disrupts the SUZ12 binding to the p15INK4B locus, increases the expression of p15INK4B, but not p16INK4A or p14ARF, and inhibits cellular proliferation. Finally, RNA immunoprecipitation demonstrates that ANRIL binds to SUZ12 in vivo. Collectively, these results suggest a model in which ANRIL binds to and recruits PRC2 to repress the expression of p15INK4B locus.

The chromodomain helicase DNA-binding (CHD) family of enzymes is thought to regulate gene expression, but their role in the regulation of specific genes has been unclear. Here we show that CHD8 is expressed at a high level during early embryogenesis and prevents apoptosis mediated by the tumour suppressor protein p53. CHD8 was found to bind to p53 and to suppress its transactivation activity. CHD8 promoted the association of p53 and histone H1, forming a trimeric complex on chromatin that was required for inhibition of p53-dependent transactivation and apoptosis. Depletion of CHD8 or histone H1 resulted in p53 activation and apoptosis. Furthermore, Chd8−/− mice died early during embryogenesis, manifesting widespread apoptosis, whereas deletion of p53 ameliorated this developmental arrest. These observations reveal a mode of p53 regulation mediated by CHD8, which may set a threshold for induction of apoptosis during early embryogenesis by counteracting p53 function through recruitment of histone H1.

HIV-1 expresses several accessory proteins to counteract host anti-viral restriction factors to facilitate viral replication and disease progression. One such protein, Vpr, has been implicated in affecting multiple cellular processes, but its mechanism remains elusive. Here we report that Vpr targets TET2 for polyubiquitylation by the VprBP-DDB1-CUL4-ROC1 E3 ligase and subsequent degradation. Genetic inactivation or Vpr-mediated degradation of TET2 enhances HIV-1 replication and substantially sustains expression of the pro-inflammatory cytokine interleukin-6 (IL-6). This process correlates with reduced recruitment of histone deacetylase 1 and 2 to the IL-6 promoter, thus enhancing its histone H3 acetylation level during resolution phase. Blocking IL-6 signaling reduced the ability of Vpr to enhance HIV-1 replication. We conclude that HIV-1 Vpr degrades TET2 to sustain IL-6 expression to enhance viral replication and disease progression. These results suggest that disrupting the Vpr-TET2-IL6 axis may prove clinically beneficial to reduce both viral replication and inflammation during HIV-1 infection.

A distinctive magnetic structure in which azimuthal and latitudinal angles of field vectors axe closely related to each other has been found in the interplanetary magnetic field data obtained by Sakigake at 0.8-1.0 AU. In this structure, termed a "planar magnetic structure" (PMS), the magnetic field vectors are nearly parallel to a fixed plane. This plane includes the spiral direction but is inclined to the ecliptic plane from 30 ø to 85 ø . The field vectors take almost all directions parallel to this plane. The PMS consists of several segments in which field directions are almost constant, and the segments axe separated by tangential discontinuities where directional changes of the field vector occur abruptly without showing any preferred polaxization. The ion number density, the ion temperature, and the plasma/3 tend to be higher in the PMS than in the surrounding plasma. The PMS events are cleaxly distinct from "magnetic clouds" both in the field configuration and in the plasma conditions. During the 25-month period from July 1985 to July 1987, eight PMS events with durations of several hours have been identified. The PMS events may be associated with newly emerging magnetic structure in the photosphere from which magnetic tongues are extended into interplanetaxy space. 1. netic field quite well [e.g., Behannon, 1978]. According to this model, the lines of force of the interplanetary magnetic field are parallel to an Archimedean spiral, and the north-south component of the field Bz is zero. While Parker's model can represent the average magnetic field of the solar wind, high time resolution data of the interplanetary magnetic field do not always agree with the model. Sometimes deviations of the field direction from the Archimedean spiral direction are as large as 90 ø , and the magnetic field vectors are almost perpendicular to the ecliptic plane. This raises a question about the causes of the deviation of the interplanetary magnetic field from the Archimedean spiral. Compression of the solar wind plasma associated with the steepening of high-speed streams [Rosenberg and Coleman, 1980] and draping of the field lines around a fast-moving plasmoid [Gosling and McComas, 1987; McComas et al., 1988] can generate directional changes of the magnetic field. However, nonspiral magnetic fields are not always associated with fast streams. Nonspiral fields which are not associated with fast streams can be due to transverse waves propagating in the interplanetary magnetic field. Alfv•n waves, which are often observed in interplanetary space, cause directional changes in the magnetic field [e.g., Belcher and Davis, 1971]. However, the deviations in the interplanetary magnetic field directions caused by Alfv•n waves would be limited to a finite angular range which is less than 90 ø with respect to the spiral direction even when the amplitude of the Alfv•n wave is large. Another candidate which can cause significant deviations from the spiral field observed in interplanetary space is the so-called "magnetic cloud" emitted fr...

SUMMARY DNA methylation at the C-5 position of cytosine (5mC) regulates gene expression and plays pivotal roles in various biological processes. The TET dioxygenases iterative oxidation of 5mC, leading to eventual demethylation intermediate. Inactivation of TET enzymes causes multi-stage developmental defects, impaired cell reprogramming and hematopoietic malignancies. However, little is known about how TET activity is regulated. Here we show that all three TET proteins bind to VprBP and are monoubiquitylated by the VprBP-DDB1-CUL4-ROC1 E3 ubiquitin ligase (CRL4VprBP) on a highly conserved lysine residue. Deletion of VprBP in oocytes abrogated paternal DNA hydroxymethylation in zygotes. VprBP-mediated monoubiquitylation promotes TET binding to chromatin. Multiple recurrent TET2-inactivating mutations derived from leukemia target either the monoubiquitylation site (K1299) or residues essential for VprBP binding. Cumulatively, our data demonstrate that CRL4VprBP is a critical regulator of TET dioxygenases during development and in tumor suppression.

SUMMARY CUL4B, encoding a scaffold protein for the assembly of Cullin4B-Ring ubiquitin ligase (CRL4B) complexes, is frequently mutated in X-linked mental retardation (XLMR) patients. Here, we show that CUL4B, but not its paralogue CUL4A, targets WDR5, a core subunit of histone H3 lysine 4 (H3K4) methyltransferase complexes, for ubiquitination and degradation in the nucleus. Knocking down CUL4B increases WDR5 and trimethylated H3K4 (H3K4me3) on the neuronal gene promoters and induces their expression. Furthermore, CUL4B depletion suppresses neurite outgrowth of PC12 neuroendocrine cells which can be rescued by co-depletion of WDR5. XLMR-linked mutations destabilize CUL4B and impair its ability to support neurite outgrowth of PC12 cells. Our results identify WDR5 as a critical substrate of CUL4B in regulating neuronal gene expression and suggest epigenetic change as a common pathogenic mechanism for CUL4B-associated XLMR.

The aim of the present study was to analyze the neuromodulation of rectoanal reflex activity by lumbar sympathetic nerves in guinea pigs. The mechanical activities of the rectum were recorded with a balloon connected to a pressure transducer, and those of the internal anal sphincter (IAS) were recorded with a custom-made strain gauge force transducer. Gradual and sustained rectal distension evoked the rectoanal reflex, causing cholinergic contractions of the rectum and synchronous nitrergic relaxations of the IAS. Section of the lumbar colonic nerves enhanced both rectal contractions and IAS relaxations. Section of the 13th thoracic cord abolished both rectal contractions and IAS relaxations, but section of the lumbar colonic nerves restored them. Lumbar sympathectomy and pithing sacral cords greatly diminished these rectal contractions and IAS relaxations, but the intrinsic reflex component remained. N G -nitro-L-arginine methyl ester enhanced the intrinsic reflex-mediated contraction of the rectum and abolished reflex-mediated relaxation of the IAS and converted into cholinergic contractions. The present results indicate that the extrinsic lumbar inhibitory outflow causes marked inhibition of the rectoanal reflex via the lumbar colonic nerves. extrinsic reflex; internal anal sphincter; intrinsic reflex; pelvic nerves; rectum WE HAVE PREVIOUSLY REPORTED (22) that a rectorectal reflex is induced by prompt rectal distension in the guinea pig. This rectorectal reflex is composed of the extrinsic excitatory reflex via sacral cords (S1-3), the extrinsic inhibitory reflex via lumbar cords (L1-4), and the intrinsic cholinergic excitatory reflex via the enteric nervous system (22). The afferent and efferent limbs of the extrinsic excitatory reflex travel in the pelvic nerves, whereas the limbs of the extrinsic inhibitory reflex pass in the lumbar colonic nerves (LCNs) (22). Furthermore, we have found that the inhibitory reflex is suppressed by descending input from the pontine defecation center, leading to a disinhibition of the sacral excitatory reflex and intrinsic excitatory reflex (22)(23)(24). In view of these findings, we have proposed that the lumbar colonic inhibitory reflex contributes to the rectorectal reflex, one important component of the defecation reflex (22-25).To clarify the integrative control of the defecation reflex by the lumbar sympathetic nerves, the goal of the present study was to elucidate the rectoanal reflex [especially the rectointernal anal sphincter (recto-IAS) reflex], because the act of defecation is a consequence of successive phenomena occurring in both the colon and anorectum (9). There is considerable evidence to support the view that the descending inhibitory reflex involving inhibitory motor neurons occurs along the entire large intestine (1, 2, 6, 7). In the current study, the rectoanal reflex (especially the recto-IAS reflex) was analyzed, and the role of the lumbar sympathetic nerves in integrative control of the distension-induced rectoanal reflex was evaluated. METHODS AND MA...

Lap-IGFI can noninvasively provide detailed lymph and blood flow information and is a useful device to aid in the accurate identification of individual patients' lymph drainage. This helps dictate adequate lymphadenectomy and the extent of intestinal resection in Lap-CRC surgery.