Disease relevance of SP6

• It is shown in this study, by means of an in vitro splicing system, that nuclear extracts obtained from cells infected with HIV-1 contain a factor (or factors) that specifically inhibits splicing of a synthetic SP6/HIV pre-messenger RNA (pre-mRNA)-containing donor and acceptor splice sites in the coding region for the envelope protein [1].
• It is also shown that the SP6/HIV pre-mRNA is not capable of assembly in a ribonucleoprotein complex, spliceosome, in extracts from infected cells [1].
• As an example, the sequence analysis is presented of M13 phage DNA and of RNA prepared by transcription with SP6 RNA polymerase [2].
• An SP6 RNA containing the adenovirus 5 L3 poly(A) site is processed efficiently in a HeLa cell nuclear extract to generate correct 3' termini [3].
• In contrast, bacteriophage SP6 RNA polymerase (RNAP) efficiently transcribes through the same nucleosome under physiological conditions, and the histone octamer is transferred behind SP6 RNAP [4].

High impact information on SP6

• A nucleosome core was assembled on a short linear DNA template (227 bp) containing an SP6 RNA polymerase promoter and a nucleosome-positioning sequence [5].
• RNA transcripts encoding the complete human glucose transporter (GT) or fragments of GT corresponding to the NH2-terminal 340 amino acids (GT-N) or the COOH-terminal 148 amino acids (GT-C) were synthesized in vitro from SP6 plasmids [6].
• We have expressed the human EGF receptor (hEGF-R) in Xenopus oocytes by injecting mRNA synthesized in vitro using SP6 vectors containing receptor cDNAs [7].
• The resulting transient heterokaryons were analyzed by using an anti-HLA-DR monoclonal antibody to assess the cell surface expression of HLA-DR (the major subtype of class II antigens) by immunofluorescence microscopy and by using uniformly 32P-labeled SP6 RNA probes in Northern blots and RNase protection assays to assess mRNA synthesis [8].
• A labeled SP6 transcript of one of the true U1 genes (pD2) was able to protect a subset of the 3' flanking sequences present in HeLa cytoplasmic U1 RNA [9].

Chemical compound and disease context of SP6

• Thymidine kinase gene (herpes simplex virus type 1) transcripts prepared in vitro using the SP6 RNA polymerase transcription system were capped and methylated posttranscriptionally with [alpha-32P]GTP and S-adenosyl-L-methionine to yield cap-labeled transcripts [10].
• Subgenomic fragments of cDNA from poliovirus type 1 were inserted downstream from the SP6 or the T7 promoter in a Gemini riboprobe vector and their in vitro synthesized RNA transcripts were used as radiolabeled probes for the detection of enteroviral RNAs by molecular hybridization [11].
• In this study we devised alternative methods to enumerate intracellular Salmonellae using a lytic bacteriophage, SP6, and an amino acid auxotroph, Pro- mutant, which replicates selectively within host cells in the presence of its uptake inhibitor, 3,4-dehydro-L-proline [12].
• When using a 5' untranslated region including SP6 promoter and Omega29 enhancer (a part of tobacco mosaic virus Omega), an A(8) sequence (eight consecutive adenylate residues) at the 3' end is suitable for fusion formation, while an XA(8) sequence (XhoI and the A(8) sequence) is suitable for C-terminal protein labeling [13].

Biological context of SP6

• The mouse Klf14 sequence is homologous to a human genomic sequence from chromosome 17 that is believed to code for a protein with three zinc fingers at the end of its C-terminal domain [14].
• A nucleosome core was assembled on a short DNA fragment and ligated into a plasmid containing a promoter and terminators for SP6 RNA polymerase [15].
• Incubation of SP6-generated pre-mRNA containing the adenovirus L3 polyadenylation site in HeLa cell nuclear extract results in the rapid assembly of specific complexes [16].
• SP6-transcribed 32P-labeled U2 small nuclear RNA precursor molecules were introduced into cultured human 293 cells by calcium phosphate-mediated uptake, as in standard DNA transfection experiments [17].
• Nuclear transport of proteins translated in vitro from SP6 plasmid-generated mRNAs [18].

Anatomical context of SP6

• In addition, SP6 IR in the outer molecular layer of the dentate gyrus was strongly correlated with tangle count in the entorhinal cortex (r = -0.70, P < 0.002), suggesting loss of perforant pathway projection [19].
• The cDNAs were expressed in vitro by SP6 polymerase transcription and rabbit reticulocyte lysate translation, and the radiolabeled protein products were analyzed by high-resolution two-dimensional gel electrophoresis [20].
• Translation of SP6 transcripts of the cDNA or of native mRNA from either normal or infected hamster brain in the absence of dog pancreas membranes resulted in the synthesis of a single PrP immunoreactive polypeptide (each polypeptide was the same size; Mr, 28,000), as predicted from the known sequence of the coding region [21].
• A novel Ets protein was isolated by yeast one-hybrid screening of a cDNA library made from lipopolysaccharide-stimulated mouse splenic B cells, using the SP6 kappa promoter kappaY element as a bait [22].
• The amino terminus of the precursor obtained by in vitro translation of SP6 alpha-chain mRNA in the presence of microsomes was leucine 23 [23].

Associations of SP6 with chemical compounds

• Incubation of SP6 generated mouse histone H4 mRNA precursors in nuclear extracts of HeLa cells yields processed mRNA species which end on the 3' adenosine of the conserved terminal ACCA sequence not unlike ten different histone mRNAs isolated from sea urchin embryos which end either on the 3' C or A [24].
• A DNA restriction fragment with convergent SP6 and T7 promoters has undergone reaction with cis-diamminedichloroplatinum(II) (cis-DDP) and was then used as a template for RNA synthesis in vitro [25].
• The ribonucleoprotein (RNP) structures of the pre-mRNA and RNA processing products generated during in vitro splicing of an SP6/beta-globin pre-mRNA were characterized by sucrose gradient sedimentation analysis [26].
• The lysate could initiate translation at methionine 1 or methionine 13, depending on the SP6 mRNA provided [27].
• Subunit-specific DNA probes for the chicken muscle acetylcholine receptor have been used in conjunction with Northern blots and dot-blots based upon the SP6/T7 RNA polymerase reaction to quantitate changes in the steady-state mRNA levels of all four subunits during development [28].

Physical interactions of SP6

• Purification and characterization of a protein binding to the SP6 kappa promoter. A potential role for CArG-box binding factor-A in kappa transcription [29].

Other interactions of SP6

• Identification of KLF13 and KLF14 (SP6), novel members of the SP/XKLF transcription factor family [14].
• SP3 (135-157 msec) and SP6 (282-339 msec) are probably generated by neurons involved in noxious somatosensation [30].
• SP6 RNAase mapping of RNA from the fused cells showed activation of the human CAI gene [31].
• SP4, SP5, and SP6 could be distinguished by changes in the orientation of the isovoltage contour lines and/or by changes in the location of the maximum [32].
• The translational efficiency of this EGF receptor RNA was found to be relatively low: approx. 100-fold lower than globin RNA synthesized using SP6 RNA polymerase [33].

Analytical, diagnostic and therapeutic context of SP6

• Transcribing SP6 RNA polymerase was arrested at unique positions in the nucleosome core, and the complexes were analyzed using biochemical methods and electron cryomicroscopy [34].
• Gel retardation assays with RNA from both wild-type and mutant TAR constructs generated in vitro with SP6 polymerase indicated specific binding of HeLa nuclear proteins to the TAR [35].
• Splicing of the regulated third intron of the L1 ribosomal protein gene of Xenopus laevis has been studied in vivo by oocyte microinjection of wild-type and mutant SP6 precursor RNAs and in vitro in the heterologous HeLa nuclear extract [36].
• SP6 analysis of the RNA isolated from the transduced cells showed that the normal beta-globin gene was transcribed at a moderately high level, before or after treatment with hemin [37].
• The adsorption and chiral expression of 6-nitrospiropyran (SP6) molecules on a Au(111) surface are studied by scanning tunneling microscopy (STM) in combination with density functional theory (DFT) calculations [38].

References