The neuronal-specific Elav-like proteins (HuD, Hel-N, and HuC) contain three RNP-type concensus motifs bind to AU-rich elements. We have identified cloned a fourth member of this family (HuR) that is expressed in wide variety cell types. purified recombinant protein binds avidly the element c-fos interleukin-3 mRNAs. In case element, HuR core 27 nucleotides AUUUA, AUUUUA, AUUUUUA motifs. Mutational analysis has shown all AU are required for maximal binding.

Expression of the cyclin-dependent kinase inhibitor p21 is highly induced by many stresses, including exposure to short-wavelength UV light (UVC), which increases mRNA stability. Investigation into mechanisms underlying this stabilization process revealed that proteins present in cytoplasmic lysates human RKO colorectal carcinoma cells formed complexes with were inducible treatment UVC and other stress agents. The ubiquitous Elav-type RNA-binding protein HuR was identified within...

Abstract We have utilized immunochemical techniques to investigate the developmental expression of Hu proteins, a neuron‐specific family RNA binding proteins in vertebrates. Previous work suggests that these may play an important role neuronal development and maintenance. For present study, we developed monoclonal antibody (MAb 16A11) binds specifically epitope gene products all known genes, including HuD, HuC, Hel‐N1. Using brief pulses (1–2 h) DNA precursor analog bromodeoxyruridine (BrdU)...

Micro-RNAs are small noncoding RNAs, which diminish the stability and/or translation of mRNAs. This study examined whether miR-206, previously shown to be elevated in estrogen receptor (ER)α-negative breast cancer, regulates expression ERα. Two putative miR-206 sites, (hERα1 and hERα2), were found silico within 3′-untranslated region human ERα mRNA. Transfection MCF-7 cells with pre-miR-206 or 2′-O-methyl antagomiR-206 specifically decreased increased, respectively, mRNA levels....

Abstract The serum and cerebrospinal fluid (CSF) of 8 women with ataxia, 6 whom also had eye movement abnormalities believed to be opsoclonus, were found contain a highly specific antineuronal antibody we call anti‐Ri. Seven the or developed cancer: carcinoma breast in 5, adenocarcinoma an axillary lymph node 1, fallopian tube 1. Four patients presented neurological disorder; cancer was diagnosed first other 4. Immunohistochemical studies using CSF from all revealed interaction central...

Abstract We looked for the presence of anti‐Hu antibody in sera from 50 normal subjects; 44 patients with small cell lung cancer, not associated paraneoplastic disease; and 25 cancer sensory neuropathy, encephalomyelitis, or both. Using avidin‐biotin immunoperoxidase method a highly sensitive quantitative Western blot analysis, was detected human sera. Seven but no syndrome had detectable levels (average titer, 76 U/ml) on blot. These are significantly lower than average titer who neuropathy...

Paraneoplastic cerebellar degeneration is a rare syndrome that occurs in patients with gynecologic cancer and characterized by widespread loss of Purkinje cells. To determine whether Purkinje-cell antigens are selectively expressed the tumors syndrome, we examined tumor tissue from 10 whose serum contained anti—Purkinje-cell (anti-Yo) antibodies. The origins cancers were breast (five patients), ovary (three), endometrium (one), fallopian tube (one). We used as controls 11 ovarian who...

The implantation process is complex, requiring reciprocal interactions between implantation-competent blastocysts and the receptive uterus. Because microRNAs (miRNAs) have major roles in regulating gene expression, we speculated that they participate directing highly regulated spatiotemporally expressed genetic network during implantation. Here, show two miRNAs, mmu-miR-101a mmu-miR-199a*, are mouse uterus coincident with expression of cyclooxygenase-2, a critical for More interestingly, our...

Human single-stranded DNA binding protein (human SSB) is a multisubunit containing polypeptides of 70, 34, and 11 kDa that required for SV40 replication in vitro. In this report we identify the functions SSB its individual subunits replication. The 70 subunit was found to bind DNA, whereas other did not. Four monoclonal antibodies against human were isolated which inhibited have been designated alpha SSB70A, SSB70B, SSB70C, SSB34A indicate are recognized. Immunolocalization experiments...

We studied the nervous systems and tumors of five patients with anti-Hu-positive paraneoplastic encephalomyelitis/sensory neuronopathy (PEM/PSN) to determine if autoantibody found in serum CSF was also present those tissues. Immunohistochemical studies system revealed presence IgG bound predominantly nuclei most neurons cytoplasm some glial cells. a lesser degree neuropil. In brains who died cancer without syndrome, immediate perivascular areas very limited There no neurons, only controls...

We measured the activity of 2 highly specific autoantibodies (anti-Yo associated with paraneoplastic cerebellar degeneration and anti-Hu sensory neuronopathy-encephalomyelitis) in simultaneously procured samples serum CSF 18 patients. also effect plasma exchange on autoantibody 6 In 11 patients degeneration, CSF/serum ratio anti-Yo antibody varied between 0.74 63, 8 having ratios substantially greater than 1. 7 neuronopathy-encephalomyelitis, from 0.6 to 44, Plasmapheresis reduced level...

Abstract The avian forebrain exhibits neurogenesis in adulthood, with neuronal production from ependymal/subependymal zone (SZ) precursor cells. To follow the commitment of newborn cells to lineage, we used their expression Hu family RNA‐binding proteins identify them before migration SZ. Adult canaries were injected [ 3 H]thymidine as a marker DNA replication, sacrificed after varying intervals, stained for Hu, and autoradiographed. We found that was not expressed by premitotic cells, but...

Abstract HuR is a ubiquitously expressed mRNA-binding protein. Intracellular localization of predominantly nuclear, but it shuttles between the nucleus and cytoplasm. In cytoplasm can stabilize certain transcripts. Because nucleocytoplasmic translocation necessary for its activity, was hypothesized that cytoplasmic expression in cancer cells could be prognostic marker. To test significance carcinogenesis breast, we have investigated mouse mammary gland tumor model from 133 invasive ductal...

The Elav-like proteins are specific mRNA binding which required for cellular differentiation. They contain three characteristic RNP2/RNP1-type RNA motifs. Previously we have shown that the first and second domains bind to AU-rich elements in 3′-UTR of mRNA. In this paper show exhibit poly(A) activity. This activity is distinct from activities been previously described. specifically long chain tails. We third domain encompasses Using poly(A)-Sepharose beads a 'sandwich' assay can...

Angiogenesis, or new blood vessel formation, is critical for the growth and spread of tumors. Multiple phases this process, namely, migration, proliferation, morphogenesis, vascular stabilization, are needed optimal tumor beyond a diffusion-limited size. The sphingosine 1–phosphate (S1P) receptor-1 (S1P1) required stabilization nascent vessels during embryonic development. Here we show that S1P1 expression strongly induced in vessels. We developed multiplex RNA interference technique to...

The RNA-binding protein HuR (also known as ELAV1) binds to the 3′-untranslated region of mRNAs and regulates transcript stability translation. However, in vivo functions are not well understood. Here, we report that murine is essential for life; postnatal global deletion Elavl1 induced atrophy hematopoietic organs, extensive loss intestinal villi, obstructive enterocolitis, lethality within 10 days. Upon deletion, progenitor cells BM, thymus, intestine underwent apoptosis, whereas quiescent...

Angiogenesis, or new blood vessel formation, is critical for the growth and spread of tumors. Multiple phases this process, namely, migration, proliferation, morphogenesis, vascular stabilization, are needed optimal tumor beyond a diffusion-limited size. The sphingosine 1–phosphate (S1P) receptor-1 (S1P1) required stabilization nascent vessels during embryonic development. Here we show that S1P1 expression strongly induced in vessels. We developed multiplex RNA interference technique to...

Cytokine stimulation of human DLD-1 cells resulted in a marked expression nitric-oxide synthase (NOS) II mRNA and protein accompanied by only moderate increase transcriptional activity. Also, there was basal transcription the NOS gene, which did not result measurable expression. The 3′-untranslated region (3′-UTR) contains four AUUUA motifs one AUUUUA motif, known to destabilize mRNAs proto-oncogenes, nuclear factors, cytokines. Luciferase reporter gene constructs containing 3′-UTR showed...

HuD is a human neuronal specific RNA-binding protein. In this study we have purified and examined its RNA binding properties in detail. binds to mRNAs that contain an AU-rich element with high affinity. the case of c-fos element, 27-nucleotide core comprising AUUUA, AUUUUA, AUUUUUA motifs. Mutation any two these motifs abrogates binding. contains tandem recognition (RRM), basic domain, third RRM. Deletion analysis has shown only first second RRMs are essential for Thus, support idea...