In vitro processing of a plant pre-mRNA in a HeLa cell nuclear extract. 1986

K Hartmuth, and A Barta

In order to determine whether there is a general difference in the splicing mechanism of animal and plant pre-mRNAs, we cloned part of the gene for the small subunit of the ribulose 1,5-bisphosphate carboxylase containing both introns into the SP64 vector. RNA was synthesized with SP6 polymerase and used as substrate for in vitro processing in a HeLa cell nuclear splicing extract. Analyses of the processed RNA demonstrate that both introns of the plant pre-mRNA are efficiently removed in an ordered fashion yielding a faithfully ligated mRNA. Two branch points were identified for intron A and three for intron B. The branched nucleotides are adenosine residues in all cases and are located within a distance from the 3' splice site found to be crucial for lariat formation in animal pre-mRNAs. The implications of these results are discussed in light of our previous observation, that a functional pre-mRNA of the human growth hormone gene was not processed in plant tissue in vivo.

UI MeSH Term Description Entries
D007438 Introns Sequences of DNA in the genes that are located between the EXONS. They are transcribed along with the exons but are removed from the primary gene transcript by RNA SPLICING to leave mature RNA. Some introns code for separate genes. Intervening Sequences,Sequences, Intervening,Intervening Sequence,Intron,Sequence, Intervening
D009694 Nucleic Acid Precursors Use for nucleic acid precursors in general or for which there is no specific heading. Acid Precursors, Nucleic,Precursors, Nucleic Acid
D010944 Plants Multicellular, eukaryotic life forms of kingdom Plantae. Plants acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations. It is a non-taxonomical term most often referring to LAND PLANTS. In broad sense it includes RHODOPHYTA and GLAUCOPHYTA along with VIRIDIPLANTAE. Plant
D002467 Cell Nucleus Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed) Cell Nuclei,Nuclei, Cell,Nucleus, Cell
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D005091 Exons The parts of a transcript of a split GENE remaining after the INTRONS are removed. They are spliced together to become a MESSENGER RNA or other functional RNA. Mini-Exon,Exon,Mini Exon,Mini-Exons
D006367 HeLa Cells The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for, among other things, VIRUS CULTIVATION and PRECLINICAL DRUG EVALUATION assays. Cell, HeLa,Cells, HeLa,HeLa Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012273 Ribulose-Bisphosphate Carboxylase A carboxy-lyase that plays a key role in photosynthetic carbon assimilation in the CALVIN-BENSON CYCLE by catalyzing the formation of 3-phosphoglycerate from ribulose 1,5-biphosphate and CARBON DIOXIDE. It can also utilize OXYGEN as a substrate to catalyze the synthesis of 2-phosphoglycolate and 3-phosphoglycerate in a process referred to as photorespiration. Carboxydismutase,Ribulose Biphosphate Carboxylase-Oxygenase,Ribulose Diphosphate Carboxylase,Ribulosebiphosphate Carboxylase,Rubisco,1,5-Biphosphate Carboxylase-Oxygenase,Ribulose Biphosphate Carboxylase,Ribulose Bisphosphate Carboxylase,Ribulose-1,5-Biphosphate Carboxylase,Ribulose-1,5-Biphosphate Carboxylase-Oxygenase,Ribulose-1,5-Bisphosphate Carboxylase Small-Subunit,Ribulose-Bisphosphate Carboxylase Large Subunit,Ribulose-Bisphosphate Carboxylase Small Subunit,Rubisco Small Subunit,1,5 Biphosphate Carboxylase Oxygenase,Biphosphate Carboxylase-Oxygenase, Ribulose,Carboxylase Small-Subunit, Ribulose-1,5-Bisphosphate,Carboxylase, Ribulose Bisphosphate,Carboxylase, Ribulose Diphosphate,Carboxylase, Ribulose-1,5-Biphosphate,Carboxylase, Ribulose-Bisphosphate,Carboxylase, Ribulosebiphosphate,Carboxylase-Oxygenase, 1,5-Biphosphate,Carboxylase-Oxygenase, Ribulose Biphosphate,Carboxylase-Oxygenase, Ribulose-1,5-Biphosphate,Diphosphate Carboxylase, Ribulose,Ribulose 1,5 Biphosphate Carboxylase,Ribulose 1,5 Biphosphate Carboxylase Oxygenase,Ribulose 1,5 Bisphosphate Carboxylase Small Subunit,Ribulose Biphosphate Carboxylase Oxygenase,Ribulose Bisphosphate Carboxylase Large Subunit,Ribulose Bisphosphate Carboxylase Small Subunit,Small Subunit, Rubisco,Small-Subunit, Ribulose-1,5-Bisphosphate Carboxylase
D012322 RNA Precursors RNA transcripts of the DNA that are in some unfinished stage of post-transcriptional processing (RNA PROCESSING, POST-TRANSCRIPTIONAL) required for function. RNA precursors may undergo several steps of RNA SPLICING during which the phosphodiester bonds at exon-intron boundaries are cleaved and the introns are excised. Consequently a new bond is formed between the ends of the exons. Resulting mature RNAs can then be used; for example, mature mRNA (RNA, MESSENGER) is used as a template for protein production. Precursor RNA,Primary RNA Transcript,RNA, Messenger, Precursors,RNA, Ribosomal, Precursors,RNA, Small Nuclear, Precursors,RNA, Transfer, Precursors,Pre-mRNA,Pre-rRNA,Pre-snRNA,Pre-tRNA,Primary Transcript, RNA,RNA Precursor,mRNA Precursor,rRNA Precursor,snRNA Precursor,tRNA Precursor,Pre mRNA,Pre rRNA,Pre snRNA,Pre tRNA,Precursor, RNA,Precursor, mRNA,Precursor, rRNA,Precursor, snRNA,Precursor, tRNA,Precursors, RNA,RNA Primary Transcript,RNA Transcript, Primary,RNA, Precursor,Transcript, Primary RNA,Transcript, RNA Primary

Related Publications

K Hartmuth, and A Barta
January 2014, Methods in molecular biology (Clifton, N.J.),
K Hartmuth, and A Barta
June 2013, Cold Spring Harbor protocols,
K Hartmuth, and A Barta
January 2014, Methods in molecular biology (Clifton, N.J.),
K Hartmuth, and A Barta
June 1989, Current opinion in cell biology,
K Hartmuth, and A Barta
June 1993, Current opinion in cell biology,
Copied contents to your clipboard!