A 22-nucleotide spliced leader sequence in the human parasitic nematode Brugia malayi is identical to the trans-spliced leader exon in Caenorhabditis elegans.

Takacs, Adrienne M.; Denker, John A.; Perrine, Kimberly G.; Maroney, Patricia A.; Nilsen, Timothy W.

doi:10.1073/pnas.85.21.7932

Cited by 53 publications

(42 citation statements)

References 15 publications

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“…Figure 1A shows the hybridization of this complementary sequence, 5'-dCTCAAACTTGGGTAATTAAACC-3', to DNAs of C. elegans and 0. volvulus restricted by three different enzymes. The probe hybridizes predominantly to a 930-bp fragment in the Avall, DdeI, and HindIII digests of C. elegans DNA, as predicted from the earlier demonstration that this sequence is encoded within the repeat unit containing the C. elegans 5S rRNA gene (28). In contrast, the probe hybridizes to more than 30 different fragments in the digests of 0. volvulus DNA.…”

Section: Resultsmentioning

confidence: 84%

“…Recently, it has been observed that 10 to 15% of the mRNAs of several nematodes, including Caenorhabditis elegans and Brugia malayi, also possess a 5' SL of 22 nucleotides whose sequence is unrelated to the 39-nucleotide SL of trypanosomes (16,28). For example, in C. elegans three of the four actin mRNA species contain this nematode SL at their 5' ends, while the fourth actin mRNA does not (16).…”

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confidence: 99%

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Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans.

Zeng¹,

Alarcon²,

Donelson³

1990

Mol. Cell. Biol.

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Section: Resultsmentioning

confidence: 84%

mentioning

confidence: 99%

Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans.

Zeng¹,

Alarcon²,

Donelson³

1990

Mol. Cell. Biol.

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“…This gene (BmATS) encodes an asparaginyl tRNA synthetase of B. malayi, and its mRNA was the first to be shown to be trans-spliced in this parasite [19]. To obtain a clone containing the promoter and downstream domains of the BmATS gene, the gene was identified by a BLAST search of The Institute for Genomic Research (TIGR) genomic sequence (available from the TIGR B. malayi genome project at http://www.tigr.org/tdb/e2k1/bma1/intro.shtml), using the full length mRNA sequence (Genbank Accession number J03266) as the query.…”

Section: Preparation Of Parental Constructsmentioning

confidence: 99%

“…To answer the first question, a portion of the B. malayi asparaginyl tRNA synthetase (BmATS) gene was isolated and tested for its ability to direct trans-splicing. The BmATS gene encodes the first mRNA shown to be trans-spliced in B. malayi [19] and it contains a perfect homologue of the BmHSP70 TSM in its first intron (Figure 4, panel A). A construct (BmATS E1-I1-E2-luc) containing the 544 nt upstream of the ORF, the first exon, the first intron and 11 nt of the second intron ( Figure 4, Panel A) was cloned upstream from the luciferase reporter gene of pGL3 basic and transfected into B. malayi embryos.…”

Section: Activity Of the Bmhsp70 Tsm In Other Genesmentioning

confidence: 99%

Sequences necessary for trans-splicing in transiently transfected Brugia malayi

Liu

Oliveira

Higazi

et al. 2007

Molecular and Biochemical Parasitology

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Many genes in parasitic nematodes are both cis-and trans-spliced. Previous studies have demonstrated that a 7nt element encoded in the first intron of the B. malayi 70 kDa heat shock protein (BmHSP70) gene was necessary to permit trans-splicing of transgenic mRNAs in embryos transfected with constructs encoding portions of the BmHSP70 gene. Here we demonstrate that this element (the B. malayi HSP70 trans-splicing motif, or BmHSP70 TSM) is necessary and sufficient to direct trans-splicing of transgenic mRNAs derived from two genes naturally containing this motif. Mutations introduced into any position of the BmHSP70 TSM abrogated its ability to direct trans-splicing. Transgenic mRNAs derived from embryos transfected with constructs containing promoters and associated downstream domains from two normally trans-spliced genes that lack a BmHSP70 TSM homologue (the B. malayi 12 kDa small subunit ribosomal protein (BmRPS12) gene and the B. malayi RNA binding protein (BmRBP1) gene), were not trans-spliced. Transfer of the BmHSP70 TSM into the first intron of the BmRPS12 gene rendered it competent for trans-splicing. Insertion of the BmHSP70 TSM into the single intron of the BmRBP1 gene did not render it transsplicing competent. However, tagged constructs of the full-length BmRBP1 gene were trans-splicing competent. An analysis of the first exons and introns of over 200 trans-spliced B. malayi genes found homologues for the BmHSP70 TSM in roughly 25%. Thus, while the BmHSP70 TSM is necessary and sufficient to direct trans-splicing in some genomic contexts, independent trans-splicing signals are employed by other genes.

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“…The C. elegans SL1 sequence is encoded within the spacer region of the repetitive DNA element containing the 5S rRNA gene cluster and transcribed from the complementary strand to an RNA of about 100 nucleotides by RNA polymerase II during early embryonic development, just like U1 snRNA molecules in vertebrates. Linkage between the 5S rRNA gene and the SL1 RNA exon within a repetitive DNA element has also been found in other nematodes such as Onchocerca volvulus [17], Dirofilaria immitis [17], Brugia malayi [13], and Ascaris lumbricoides [14]. Although the 5S rRNA and SL1 loci in C. elegans are on complementary strands of the repetitive DNA element, the two loci for 5S rRNA and SL1 are on the same strand in the parasitic nematodes, O. volvulus, D. immitis, B. malayi, and A. lumbricoides.…”

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confidence: 99%

Characteristics of Nucleotide Sequences Flanking the trans-Spliced Leader SL1 Exon in Dirofilaria immitis, Brugia malayi, and Brugia pahangi.

Harasawa

Maeda

Nogami

et al. 1997

The Journal of Veterinary Medical Science

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ABSTRACT. Nucleotide sequences surrounding the trans-spliced leader SL1 exon in the 5S rRNA gene spacer regions of Dirofilaria immitis, Brugia malayi, and B. pahangi were determined after PCR amplification, aligned with the genus Onchocerca for comparison, and used for the prediction of secondary structures. The nucleotide sequence of this region in B. pahangi was first shown in the present study. Hypothetical secondary structures of the spacer region suggested that the SL1 transcript is capable to form a stable stem-loop structure which may render transposition of the SL1 sequence to mRNA molecules. A homologous sequence to Sm-binding site was assigned on a bulge loop. No significant difference was observed in adult worms of D. immitis irrespective of sex or location. No difference was apparent between the two species in genus Brugia. -KEY WORDS: Brugia, Dirofilaria, trans-splicing. J. Vet. Med. Sci. 59(12): 1149-1152, 1997 Of the four adult worms of D. immitis, two (male and female, each) were from Japan, which are referred to as "Japanese adult worms", and two (male and female, each) were from the U.S.A. which are referred to as "American adult worms", hereafter. Japanese adult worms in the heart were collected from naturally infected dogs at necropsy [11]. American adult worms were collected from naturally infected dogs. To determine the prevalence of D. immitis infection in dogs, these dogs were killed by euthanasic procedure and immediately provided from a dog pound of Wake County, Raleigh, North Carolina, U.S.A. Two Brugia species, B. malayi and B. pahangi, have been maintained in the second author's laboratory. Exact source of the B. malayi strain has been described elsewhere [9]. All adult worms were identified by morphological features. DNA of D. immitis was isolated from a single adult worm, and DNA of Brugia was extracted from frozen specimens of adult worms by the hydroxyapatite batch elution technique [3]. Parasite DNAs were subjected to PCR to amplify the 5S rRNA gene and its 3' flanking spacer region by using a pair of primers based on genomic sequences of high homology between Dirofilaria and Onchocerca [17]. The PCR primers F1 (5'-GTC TAC GAC CAT ACC ACG TT-3') and R1 (5'-CAG GAC GTT CCA AAA TTT-3') were custom-made by Takara Shuzo Co., Ltd. (Kyoto). The PCR amplification was carried out with 50-µl reaction mixtures containing 5 µl of DNA solution, 5 µl of 10X buffer, 1.25 U of Taq DNA polymerase (Perkin-Elmer Corp., Norwalk, Conn.), dNTPs to a final concentration of 0.2 mM each, 0.25 µl of each pair of primers F1 and R1 (40 pmol/µl each), and water to a final volume of 50 µl. The cycle was repeated 30 times with denaturation at 94°C for 30 sec, annealing at 56°C for 120 sec, and extension at 72°C for 120 sec. The PCR products were extracted from gels after electrophoresis, and subjected to direct sequencing three times on each strand A spliced leader sequence at the 5' end of a mRNA was first demonstrated in African trypanosomes [2]. This trypanosome leader sequence is derived from an RNA tra...

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A 22-nucleotide spliced leader sequence in the human parasitic nematode Brugia malayi is identical to the trans-spliced leader exon in Caenorhabditis elegans.

Cited by 53 publications

References 15 publications

Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans.

Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans.

Sequences necessary for trans-splicing in transiently transfected Brugia malayi

Characteristics of Nucleotide Sequences Flanking the trans-Spliced Leader SL1 Exon in Dirofilaria immitis, Brugia malayi, and Brugia pahangi.

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