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Query: UNIPROT:P06889 (Mol)
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It has been proposed that recognition of the 3' splice site in many group I introns involves base pairing between the start of the 3' exon and a region of the intron known as the internal guide sequence (R. W. Davies, R. B. Waring, J. Ray, T. A. Brown, and C. Scazzocchio, Nature [London] 300:719-724, 1982). We have examined this hypothesis, using the self-splicing rRNA intron from Tetrahymena thermophila. Mutations in the 3' exon that weaken this proposed pairing increased use of a downstream cryptic 3' splice site. Compensatory mutations in the guide sequence that restore this pairing resulted in even stronger selection of the normal 3' splice site. These changes in 3' splice site usage were more pronounced in the background of a mutation (414A) which resulted in an adenine instead of a guanine being the last base of the intron. These results show that the proposed pairing (P10) plays an important role in ensuring that cryptic 3' splice sites are selected against. Surprisingly, the 414A mutation alone did not result in activation of the cryptic 3' splice site.
Mol Cell Biol 1990 Jun
PMID:Base pairing between the 3' exon and an internal guide sequence increases 3' splice site specificity in the Tetrahymena self-splicing rRNA intron. 234 65

The human cellular oncogene c-myc contains two promoters at the 5' end of its first non-coding exon. Cryptic promoters located within the first intron are also activated to synthesize aberrant c-myc mRNAs in some Burkitt lymphomas with a t(8: 14) chromosome translocation in which a part of the gene structure, often the 5' non-coding exon, is truncated. We have shown elsewhere that microinjected plasmid DNA carrying a normal, intact human c-myc gene directs efficient faithful transcription from its own two promoters in Xenopus laevis oocytes. Here, I have investigated the expression of different recombinants carrying various constructs of the c-myc gene in frog oocytes in order to understand the activation mechanism of those cryptic promoters. Aberrant c-myc transcripts initiating from cryptic promoters within the first intron are undetectable when the intact or truncated c-myc gene construct is used. However, the cryptic promoters can be activated in Xenopus oocytes if the truncated c-myc gene construct is fused with simian virus 40 sequences containing a 21 base-pair repeat and the replication origin. Xenopus oocytes will be useful for further investigation of enhancing elements involved in the translocated and activated c-myc genes in Burkitt lymphoma cells.
J Mol Biol 1987 Feb 05
PMID:Activation of cryptic promoters of human c-myc genes in microinjected Xenopus laevis oocytes. 243 22

Two murine monocytic leukemia cell lines, WEHI-265 and WEHI-274, were found to carry a rearranged c-myb gene. The rearrangements are due to insertion of a deleted Moloney murine leukemia virus (Mo-MLV) provirus in the 5' region of the c-myb gene and thus are similar to rearrangements in the ABPL tumors (G. L. C. Shen-Ong, M. Potter, J. F. Mushinski, S. Lavu, and E. P. Reddy, Science 226:1077-1080, 1984). In each cell line, the retroviral insertion has induced high levels of two aberrant RNA species, which, as in the ABPL tumors (G. L. C. Shen-Ong, H. C. Morse, M. Potter, and J. F. Mushinski, Mol. Cell. Biol. 6:380-392, 1986), contain both viral (Mo-MLV) and cellular (myb) sequences. Both species lack the sequences encoding the amino terminus of the c-myb protein and thus could encode a protein which, like the v-myb gene products (and the predicted ABPL myb proteins), is truncated at the amino terminus. We have found that the larger (5.3 kilobase [kb]) and more abundant of the tumor-specific myb RNAs was predominantly nuclear, while the smaller species (3.9 kb) was cytoplasmic. Furthermore, our data imply that the 3.9-kb RNA was derived from the 5.3-kb RNA by an additional splice which utilized a cryptic splice acceptor site within the viral gag sequences. On the basis of subcellular distribution and predicted translational potential, we conclude that the 3.9-kb RNA is probably the mRNA which encodes a truncated myb protein. We also show that, due to different insertion points in W265 and W274, the W274 myb RNAs contained sequences from a c-myb exon upstream of the exons represented in the W265 (and ABPL) RNAs. The significance of our findings with regard to transformation by myb in these tumors is discussed.
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PMID:Generation of altered transcripts by retroviral insertion within the c-myb gene in two murine monocytic leukemias. 244 Oct 77

The intervening sequence (IVS) of the Tetrahymena thermophila ribosomal RNA precursor undergoes accurate self-splicing in vitro. The work presented here examines the requirement for Tetrahymena rRNA sequences in the 5' exon for the accuracy and efficiency of splicing. Three plasmids were constructed with nine, four and two nucleotides of the natural 5' exon sequence, followed by the IVS and 26 nucleotides of the Tetrahymena 3' exon. RNA was transcribed from these plasmids in vitro and tested for self-splicing activity. The efficiency of splicing, as measured by the production of ligated exons, is reduced as the natural 5' exon sequence is replaced with plasmid sequences. Accurate splicing persists even when only four nucleotides of the natural 5' exon sequence remain. When only two nucleotides of the natural exon remain, no ligated exons are observed. As the efficiency of the normal reaction diminishes, novel RNA species are produced in increasing amounts. The novel RNA species were examined and found to be products of aberrant reactions of the precursor RNA. Two of these aberrant reactions involve auto-addition of GTP to sites six nucleotides and 52 nucleotides downstream from the 3' splice site. The former site occurs just after the sequence GGU, and may indicate the existence of a GGU-binding site within the IVS RNA. The latter site follows the sequence CUCU, which is identical with the four nucleotides preceding the 5' splice site. This observation led to a model where where the CUCU sequence in the 3' exon acts as a cryptic 5' splice site. The model predicted the existence of a circular RNA containing the first 52 nucleotides of the 3' exon. A small circular RNA was isolated and partially sequenced and found to support the model. So, a cryptic 5' splice site can function even if it is located downstream from the 3' splice site. Precursor RNA labeled at its 5' end, presumably by a GTP exchange reaction mediated by the IVS, is also described.
J Mol Biol 1987 Jul 05
PMID:5' exon requirement for self-splicing of the Tetrahymena thermophila pre-ribosomal RNA and identification of a cryptic 5' splice site in the 3' exon. 244 17

The membranes of Plasmodium falciparum-infected human red blood cells contain antigens of demonstrably cryptic character. We show here, by a cell surface radioimmunoassay using anti-human red cell membrane antisera, that raising the membrane microviscosity of intact cells leads to a marked increase in the cell surface antigen reactivity of normal cells, and even more so in cells infected in vitro with two strains of P. falciparum. A variety of sera from adults and children living in endemic areas and from malaria patients, all of which showed no detectable surface reactivity with either normal or infected red cells, were demonstrably surface-reactive to infected cells whose sterol membrane content has been raised by means conservative of cell integrity. New epitopes become exposed on the surface of infected cells after lipid modification. The present studies indicate that the reduced membrane viscosity reported in malaria-infected cells determines to a considerable extent the expression of cell surface antigens of both host and parasite, and could play a significant role in parasite immune evasion.
Mol Biochem Parasitol 1989 Sep
PMID:Passive modulation of antigenic expression in the surface of normal and malaria-infected erythrocytes. 247 77

Almost all clinical isolates of Neisseria gonorrhoeae harbour a plasmid of 4.2 kb with no known function. A genetic model based on the DNA sequence of the plasmid, with ten open reading frames, has been proposed by Korch et al., (1985). To address the question of the function of the encoded proteins, some of which are expressed when the plasmid is harboured by Escherichia coli, the subcellular locations of such proteins were investigated in minicells of Escherichia coli DS410. The protein CppB, earlier proposed to be a membrane-spanning polypeptide, was found associated with the outer membrane. Up to five other cryptic plasmid proteins were found to be localized in the periplasm.
Mol Microbiol 1989 Oct
PMID:Subcellular localization of proteins encoded by the phenotypically cryptic plasmid of Neisseria gonorrhoeae: biological evidence for outer membrane association of the cppB gene product. 251 15

We report the isolation and characterization of the mouse carbonic anhydrase I (CAI) gene. Direct RNA sequence analysis of the 5' nontranslated regions of CAI mRNA from mouse colon and mouse erythroleukemia cells demonstrated tissue specificity in the lengths and sequences of CAI transcripts. Analysis of several mouse CAI genomic clones showed that the transcripts arose from a single CAI gene with two tissue-specific promoters and eight exons. CAI transcripts in the colon were found to initiate just upstream of the erythroid exon 2 of the CAI gene region sequence. Erythroid transcripts originated from a novel promoter upstream of exon 1, which was located more than 10 but less than 250 kilobases upstream of exon 2. Erythroid exon 1 contained only a nontranslated sequence, which was spliced to exon 2 via a cryptic splice acceptor site located in the region that encoded the colon mRNA 5' nontranslated sequence. The remaining exon-intron junctions were conserved in comparison with those of the CAII and CAIII genes.
Mol Cell Biol 1989 Aug
PMID:The mouse carbonic anhydrase I gene contains two tissue-specific promoters. 257 23

The polyadenylation signal of a pea gene for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcS) has been analyzed by deletion mutagenesis and Ti plasmid-mediated gene transfer. Sequences between 6 and 137 bases upstream from the normal polyadenylation sites in this gene (bases -6 to -137) are required for functioning of these sites. In addition, bases -111 to -235 can affect 3' end formation by altering the pattern of 3' termini seen in various transcription units. Sequences between 37 and 95 bases upstream from a cryptic polyadenylation site in this gene [A. G. Hunt, DNA 7: 329-336 (1988)] are necessary for mRNA 3' end formation at this site. At least two different parts of the 3' region of this rbcS gene can serve as a downstream element for polyadenylation at the normal poly(A) addition sites in this gene. Our studies indicate that: 1. the upstream sequences required for polyadenylation in plants are different from those defined in mammalian RNA polymerase II transcription units; 2. sequences 100 or more bases upstream and downstream from poly(A) addition sites in this gene can affect poly(A) addition site choice; and 3. there are apparently redundant downstream elements for polyadenylation in this gene.
Plant Mol Biol 1989 Aug
PMID:Deletion analysis of the polyadenylation signal of a pea ribulose-1,5-bisphosphate carboxylase small-subunit gene. 257 6

Bacteriophage T4 gene 32 lies at the 3' end of a complex transcription unit which includes genes 33, 59, and several open reading frames. In the course of an infection, four major transcripts are synthesized from this unit: two overlapping polycistronic transcripts about 3800 and 2800 nucleotides in length, and two monocistronic gene 32 transcripts about 1150 and 1100 nucleotides in length. These transcripts are made at different times in infection and the polycistronic transcripts have segmental differences in stability. Messenger RNA processing yields a 1025 nucleotide monocistronic gene 32 transcript, and a 135 nucleotide transcript containing part of the gene 59 coding sequence. Processing depends on Escherichia coli encoded ribonuclease E. This pattern of transcription and processing leads to the synthesis of gene 32 mRNA throughout infection, whereas transcripts encoding the upstream genes are present only early in infection. The 3800 nucleotide polycistronic transcript initiates at a promoter that does not require T4 encoded factors for activity. However, full-length synthesis of this transcript depends on the T4 mot gene product. The region upstream of gene 32 also contains four E. coli-like promoters that are active on chimeric plasmids in uninfected cells, but inactive in bacteriophage T4. The location of these cryptic T4 promoters is intriguing in that they lie near the 5' ends of open reading frame B, gene 59 and gene 32. They could play a role in phage development under particular conditions of growth or in bacterial hosts other than those examined here.
Mol Gen Genet 1989 Oct
PMID:Transcription and messenger RNA processing upstream of bacteriophage T4 gene 32. 261 64

The areAr-18 mutation is a loss-of-function mutation in areA, the positive acting regulatory gene mediating nitrogen metabolite repression in Aspergillus nidulans. It results from a reciprocal translocation which splits the coding region into 5' and 3' moieties. Surprisingly, we have selected rare intracistronic revertants of areAr-18. From crosses heterozygous for areAr-18 revertant alleles, duplication-deficiency progeny containing two copies of a substantial portion of chromosome IV but lacking part of chromosome III, including the 5' moiety of areA, have been obtained. For all four revertants analysed genetically, growth properties of these duplication-deficiency strains indicate that the reversion events involve the 3' portion of areA and that the 5' portion of areA is unnecessary for the revertant phenotype. This conclusion was directly confirmed for one revertant using Southern blotting. As all four reversion events involve additional chromosomal rearrangements, they probably fuse functional promoters, ribosome binding sites and 'in frame' initiation codons to the 3' portion of the gene. In the course of characterisation of these mutations, new mapping data for a large region of chromosome IV have been generated, and a new reciprocal translocation activating the cryptic regulatory gene areB, whose product can substitute for that of areA, has been identified.
Mol Gen Genet 1989 Jan
PMID:A translocation associated, loss-of-function mutation in the nitrogen metabolite repression regulatory gene of Aspergillus nidulans can revert intracistronically. 265 86


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