The levels of histone mRNA increase 35-fold as selectively detached mitotic CHO cells progress from mitosis through G1 and into S phase. Using an exogenous gene with a 3' end which is not sensitive to transcriptional or half-life regulation, we show that processing regulated similar in G1- S-phase cells, measured after inhibition transcription by actinomycin D (dactinomycin) indirectly stabilization the protein synthesis inhibitor cycloheximide. Taken together, these results suggest change...

A set of seven Sm proteins assemble on the Sm-binding site spliceosomal U snRNAs to form ring-shaped core. The U7 snRNP involved in histone RNA 3′ processing contains a structurally similar but biochemically unique core which two these proteins, D1 and D2, are replaced by Lsm10 another as yet unknown component. Here we characterize this factor, termed Lsm11, novel Sm-like protein with apparently distinct functions. In vitro studies suggest that its long N-terminal part mediates an important...

Spinal muscular atrophy (SMA) is caused by reduced levels of survival motor neuron (SMN) protein, which results in motoneuron loss. Therapeutic strategies to increase SMN including drug compounds, antisense oligonucleotides, and scAAV9 gene therapy have proved effective mice. We wished determine whether reduction postnatal motoneurons resulted SMA a large animal model, could be corrected after development muscle weakness, the response clinically relevant biomarkers.Using intrathecal delivery...

In vitro-synthesized transcripts of the sea urchin histone H2A gene with 3' extensions are efficiently and rapidly processed to mRNA faithful ends in Xenopus laevis oocyte nuclei. Processing requires presence a histone-specific dyad symmetry element H2A-proximal spacer sequences precursor RNA. DNA injection experiments processing-deficient mutant, transcription products appear terminate heterogeneously first 100-200 base pairs post-H2A spacer. synthetic H3 RNA precursors prior 60-nucleotide...

We have analyzed the expression of endogenous histone H4 genes and a newly introduced gene in 21-Tb cells, mouse mastocytoma cell-cycle mutant. Endogenous mRNAs were less abundant by factor 120-180 G1-arrested than exponentially multiplying cells. However, transcription rates only decreased 3 under these conditions, as determined vitro elongation nascent transcripts. This indicates that post-transcriptional control mRNA levels is important, accord with published data. gene, modified...

In both rabbit poxvirus and vaccinia virus DNA have demonstrated an identical distribution of eight HinfI. The length the terminal repeats was found to be 3.4 3.6 megadaltons (Mdaltons) for 7.4 8.0 Mdaltons DNA. Maps HinfI restriction sites within isolated EcoRI end fragments PHAVE DEMONSTRATED AN IDENTICAL DISTRIBUTION OF EIGHT in internal part (approximately 2 Mdaltons) two genomes.

In several forms of β-thalassemia, mutations in the second intron β-globin gene create aberrant 5′ splice sites and activate a common cryptic 3′ site upstream. As result, thalassemic pre-mRNAs are spliced almost exclusively via leading to deficiency correctly mRNA and, consequently, β-globin. We have designed series vectors that express modified U7 snRNAs containing sequences antisense either or IVS2–705 pre-mRNA. Transient expression HeLa cell line stably expressing restored up 65% correct...

We have utilized a recombinant vector system designed to study gene control elements by fusing them the Escherichia coli galactokinase (galK) examine effects of expression that result from introducing changes into 5' noncoding leader region galK transcription unit. varied length and found no effect on expression, provided initiation codon remained first AUG in Using synthetic linkers, we then inserted specific sequences, each containing single at defined position within region. inserts had...

Ultrastructural analysis of the polydnavirus braconid wasp Chelonus inanitus revealed that virions consist one cylindrical nucleocapsid enveloped by a single unit membrane. Nucleocapsids have constant diameter 33·7 ± 1·4 nm and variable length between 8 46 nm. Spreading viral DNA showed genome consists circular dsDNA molecules sizes measurement contour lengths indicated 7 31 kbp. When were exposed to osmotic shock conditions release DNA, only molecule was released per particle suggesting...

In spinal muscular atrophy (SMA), the leading genetic cause of early childhood death, survival motor neuron 1 gene ( SMN1 ) is deleted or inactivated. The nearly identical SMN2 has a silent mutation that impairs utilization exon 7 and production functional protein. It been hypothesized therapies boosting inclusion might prevent cure SMA. Exon can be stimulated in cell culture by oligonucleotides intracellularly expressed RNAs, but evidence for an vivo improvement SMA symptoms lacking. Here,...

The DNAs of two closely related orthopoxviruses, rabbit poxvirus (RPV) and vaccinia virus (VV), were mapped by overlapping-fragment analysis using restriction endonucleases HindIII Sst I. exact arrangement these fragments was accomplished total digestion isolated partial products end-fragment determination. RPV VV showed identical patterns in an internal region comprising approximately 60% the genome. size, electrophoretical DNA, 118 X 10(6) daltons, some 6 daltons less than DNA. opposite...

Invariant Natural Killer T cells (iNKT) are a versatile lymphocyte subset with important roles in both host defense and immunological tolerance. They express highly conserved TCR which mediates recognition of the non-polymorphic, lipid-binding molecule CD1d. The structure human iNKT TCRs is unique that only one six complementarity determining region (CDR) loops, CDR3β, hypervariable. role this loop for biology has been controversial, it unresolved whether contributes to TCR:CD1d binding or...

Many endoparasitic wasps inject, along with the egg, polydnavirus into their insect hosts, virus being a prerequisite for successful parasitoid development. The genome of polydnaviruses consists multiple circular dsDNA molecules variable size. We show 12 kbp segment braconid Chelonus inanitus (CiV12) that it is integrated wasp genome. This first direct demonstration integration bracovirus. PCR data indicated form CiV12 was present in all male and female stages investigated while excised DNA...

Oligonucleotides derived from the spacer element of histone RNA 3' processing signal were used to characterize mouse U7 small nuclear (snRNA), i.e., snRNA component active in pre-mRNA. Under RNase H conditions, such oligonucleotides inhibited reaction, indicating formation a DNA-RNA hybrid with functional ribonucleoprotein component. Moreover, these hybridized single species approximately 65 nucleotides. The sequence this was determined by primer extension experiments and found bear several...

Spinal muscular atrophy (SMA) is a lethal hereditary disease caused by homozygous deletion/inactivation of the survival motoneuron 1 (SMN1) gene. The nearby SMN2 gene, despite its identical coding capacity, only an incomplete substitute, because single nucleotide difference impairs inclusion seventh exon in messenger RNA (mRNA). This splicing defect can be corrected (transiently) specially designed oligonucleotides. Here we have developed more permanent correction strategy based on...

We present the nucleotide sequence of galactokinase gene ( gal K) Escherichiacoli including its 5′ and 3′ flanking regions. This DNA derives from λga18 transducing phage is identical to in k fusion vectors, pKO pKG, commonly used study transcriptional regulatory elements. define precise junction between bacterial sequences demonstrate that this prohably results a homologous recombination event 9 bp common operon λ. Moreover, we examine 300 region located immediately beyond K for...