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Enzyme
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Query: EC:2.7.7.48 (
transcriptase
)
9,479
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Reverse
transcriptase
has been shown to transcribe virion DNA of influenza A virus without an exogeneous primer. At least six virion RNA segments are transcribed with the formation of complementary 4S DNA product. A possible primer function of hairpin structures at the 3'-end of virion RNA segments is discussed.
Mol
Biol (Mosk)
PMID:[Reverse transcription of influenza virion RNA without exogenous primer]. 617 95
Partially purified
RNA-dependent RNA polymerase
(RdRp) isolated from plants infected with turnip crinkle virus (TCV) is capable of template-dependent synthesis of TCV-associated RNAs. To determine the cis-sequences required for the synthesis of TCV satellite (sat-) RNA C (-) strands in vitro, templates containing interior deletions were subjected to transcription using RdRp-active fractions. Results indicated that the promoter for (-)-strand synthesis was contained within the 3'-terminal 29 bases of the (+)-strand. Structural probing by enzymatic digestion and chemical modification revealed the presence of a hairpin structure within this terminal region. Compensatory exchanges of four bases in the lower stem or alterations in the sequence and size of the loop region did not affect in vitro transcription, implying that the primary sequence in the loop and lower part of the stem is not important for interaction with the viral RdRp. However, single mutations in the base of the stem or double mutations in the upper stem strongly reduced template activity in vitro, suggesting that the stability of the hairpin is an important functional consideration. Relocation of the 3'-terminal 37 bases containing this stem-loop to inactive template RNA rendered the resultant hybrid RNA competent for in vitro transcription by RdRp activity, suggesting that the promoter for (-)-strand synthesis in vitro is completely contained within the 3'-terminal region.
J
Mol
Biol 1995 Nov 17
PMID:Requirement of a 3'-terminal stem-loop in in vitro transcription by an RNA-dependent RNA polymerase. 747 59
The gypsy group of long-terminal-repeat retrotransposons contains elements having the same order of enzyme domains in the pol gene as do retroviruses. Elements in the gypsy group are now known from yeast, filamentous fungi, plants, insects, and echinoids. Reverse
transcriptase
and RNase H amino acid sequences from elements in the gypsy group--including the recently described SURL elements, TED, Cft1, and Ulysses,--were aligned and analyzed by using parsimony and bootstrapping methods, with plant caulimoviruses and/or retroviruses as outgroups. Clades supported at the 95% level after bootstrapping include (1) 17.6 with 297 and (2) all of the SURL elements together. Other likely relationships supported at lower bootstrap confidence intervals include (1) SURL elements with mag, (2) 17.6 and 297 with TED, and this collective group with 412 and gypsy, (3) Tf1 with Cft1, (4) IFG7 with Del, and (5) all of the retrotransposons in the gypsy group together, to the exclusion of Ty3. In contrast with an earlier analysis, our results place mag within the gypsy group rather than outside of a cluster that contains gypsy group retrotransposons and plant caulimoviruses. Several features of retrotransposon genomes provide further support for some of the aforementioned relationships. The union of SURL elements with mag is supported by the presence of two RNA binding sites in the nucleocapsid protein. Location of the tRNA primer binding site and the presence of a long open reading frame 3' to the pol gene support the 17.6-297-TED-412-gypsy cluster.
Mol
Biol Evol 1993 Nov
PMID:Phylogenetic relationships of reverse transcriptase and RNase H sequences and aspects of genome structure in the gypsy group of retrotransposons. 750 45
Because surfactant proteins A, B, and C (SP-A, SP-B, and SP-C) are putative markers for alveolar epithelial type II cells, we investigated their expression in embryonic rat lung (12 to 15 days, term = 22 days). The expression of the messages for SP-A, SP-B, and SP-C was assessed by the reverse-
transcriptase
polymerase chain reaction (RT-PCR). Embryonic rat lung at 12 days' gestation lacked detectable mRNAs for all three surfactant proteins. Messages for SP-A, SP-B, and SP-C were, however, present in embryonic rat lung at 13 days' gestation. Expression of SP-A mRNA increased in embryonic rat lung with advancing gestation. Surfactant protein mRNA expression during the embryonic period of lung development was limited to the epithelial cells. The expression of SP-A mRNA, while limited to the lung, did not appear to be a marker for cells destined to become type II cells, since it was detected in the trachea and the presumptive main bronchial ducts of embryonic rat lung at 13 days' gestation, as well as in the distal lung prealveolar region. Expression of both SP-B and SP-C mRNAs in embryonic lung was confined to the distal portion of the ductal system, although message of SP-C was also found in brain and kidney. These results suggest that none of the three surfactant protein mRNAs studied are, at this early stage of lung development, specific for cells destined to become type II pneumocytes.
Am J Respir Cell
Mol
Biol 1994 Feb
PMID:Expression of surfactant proteins in embryonic rat lung. 750 64
The 3' terminus of the (-) RNA strand in the replicative forms of several (+)-stranded RNA viruses possesses an unpaired guanosine with unknown function. This unpaired guanosine is also found at the 3' terminus of the (-) strand in the double-stranded form of two cucumoviral satellite RNAs. Using a cucumber mosaic virus (CMV)
RNA-dependent RNA polymerase
capable of replicating the satellite RNA in vitro, the 3'-terminal guanosine of the satellite (-) strand was shown to be an absolute requirement for satellite (+) strand synthesis. If genomic RNA synthesis of CMV and other members of the alphavirus-like superfamily that produce (-) strands terminating in an unpaired 3' guanosine follows a similar strategy, the work reported here would represent the first experimental support for the notion of 3'-terminal guanosine functioning as an essential recognition signal for viral replicases, enabling (+) strand RNA synthesis to be initiated internally from a (-) strand template.
J
Mol
Biol 1994 May 20
PMID:Requirement of 3'-terminal guanosine in (-)-stranded RNA for in vitro replication of cucumber mosaic virus satellite RNA by viral RNA-dependent RNA polymerase. 751 30
Reverse
transcriptase
(RT) from the human immunodeficiency virus type 1 has been crystallized in four closely related forms, the best of which diffract X-rays to 2.2 A resolution. The RT was crystallized as a complex with a non-nucleoside inhibitor, either nevirapine or a nevirapine analogue. Crystals grew from 6% PEG 3400 buffered at pH 5. These were of space group P2(1)2(1)2(1) with unit cell parameters a = 147 A, b = 112 A, c = 79 A (form A), with one RT heterodimer in the asymmetric unit. Changes in unit cell parameters and degree of crystalline order were observed on soaking pregrown crystals in various solutions, giving three further sets of unit cells. These were a = 143 A, b = 112, A, c = 79 A (form B), a = 141 A, b = 111 A, c = 73 A (form C), a = 143 A, b = 117 A, c = 66.5 A (form D). The last two forms diffract X-rays to 2.2 A resolution. Structure determinations of these latter crystal forms of RT should give a detailed atomic model for this therapeutically important drug target.
J
Mol
Biol 1994 Sep 30
PMID:Crystals of HIV-1 reverse transcriptase diffracting to 2.2 A resolution. 752 79
Reverse
transcriptase
-like sequences (RTs) with amino acid motifs characteristic of non-LTR retrotransposons have been isolated from several medically important phlebotomine sandfly species. These sequences were amplified using the polymerase chain reaction (PCR) with primers based on conserved amino acid motifs present in previously described insect RTs. A further set of RTs were amplified using primers based on the conserved regions identified in phlebotomine RTs. The average similarity of the phlebotomine RTs to the Drosophila I, F and R1Dm elements was 28-29% between the closest primers used. Phlebotomine RTs were 31-62% similar to each other, the most dissimilar sequences coming from the same species. Several amino acid residues were invariant in the ten phlebotomine RTs, including motif Q(F/Y)GF, conserved in other non-LTR retrotransposons, but not in retrovirus or LTR retrotransposon RTs. The remarkable conservation of this distinctive domain of non-LTR retrotransposon RTs suggests it has a vital and possibly unique role in the mode of reverse transcription of this class of transposon.
Insect
Mol
Biol 1994 May
PMID:Isolation of non-LTR retrotransposon reverse transcriptase-like sequences from phlebotomine sandflies. 752 83
11 beta-Hydroxysteroid dehydrogenase (11 beta-HSD), responsible for the interconversion of hormonally active cortisol to inactive cortisone, dictates specificity for the mineralocorticoid receptor (MR) in the distal nephron and colon. Two isoforms of human 11 beta-HSD have been cloned, an NADP(H)-dependent (type 1) dehydrogenase/oxo-reductase enzyme, and a high-affinity NAD-dependent (type 2) unidirectional dehydrogenase. Using the reverse-
transcriptase
polymerase chain reaction (RT-PCR) amplification of RNA extracted from human adult tissues, type 1 11 beta-HSD mRNA was found in decidua, placenta, liver, lung, spleen, kidney medulla, cerebellum and pituitary, but was absent in kidney cortex, sigmoid and rectal colon, salivary gland and thyroid. In contrast, type 2 11 beta-HSD mRNA was found only in placenta and in the classical mineralocorticoid target tissues, kidney cortex, kidney medulla, sigmoid and rectal colon, salivary gland, and colonic epithelial cell lines (AAC1 and RGC28). In situ hybridization studies of renal cortex, cortico-medullary junction and medulla using a 35S-labeled antisense cRNA probe for type 2 human 11 beta-HSD, revealed specific localization of type 2 11 beta-HSD mRNA expression exclusively to renal cortical and medullary collecting ducts. Type 1 and type 2 isoforms of human 11 beta-HSD are expressed in a distinct tissue-specific fashion, in keeping with the proposed differences in their physiological roles. Type 2 11 beta-HSD is found predominantly in mineralocorticoid target tissues where it serves to protect the MR in an autocrine fashion.
Mol
Cell Endocrinol 1995 Apr 28
PMID:Detection of human 11 beta-hydroxysteroid dehydrogenase isoforms using reverse-transcriptase-polymerase chain reaction and localization of the type 2 isoform to renal collecting ducts. 754 19
In nasal biopsies from 17 adult patients with seasonal allergic rhinitis and from 10 healthy controls, cytokines were analyzed by reverse-
transcriptase
polymerase chain reaction (RT-PCR). The time-course study during winter included repeated local allergen provocation with subsequent nasal biopsies as well as biopsies taken during pollen season. The RT-PCR for CD44 yielded positive bands in 65 of 71 cases, in which cases mRNA for interleukins 2, 4, and 5 (IL-2, IL-4, and IL-5) were thus investigated by means of seminested PCR. IL-4 mRNA was found almost exclusively in the allergic patients. During provocation a significant increase in IL-4 was noticed compared with controls (p = 0.043). Equally, during the natural pollen season, IL-4 mRNA expression was significantly higher in patients not using nasal corticosteroids compared with those who did (p = 0.011). No differences in IL-2 or IL-5 were observed between the groups. These findings also indicate, together with earlier observations of T-cell activation, a phenotype switch toward T-helper 2 (Th2) cells, and the accumulation (homing) of these T cells in the nasal mucosa, that T cells constitute the main source for IL-4 in the nasal mucosa. Therefore, allergic patients have an increased synthesis of IL-4 when provoked with the allergen, and during natural pollen season this synthesis can be downregulated by corticosteroids. Furthermore, this study exemplifies the versatility of molecular biology in surgical pathology and that even low-copy-number cytokine mRNA can be examined in routinely snap-frozen surgical specimens.
Diagn
Mol
Pathol 1995 Jun
PMID:Nasal messenger RNA expression of interleukins 2, 4, and 5 in patients with allergic rhinitis. 755 Dec 98
The entire sequence of 13952 nucleotides of a plasmid-like, double-stranded RNA (dsRNA) from rice was assembled from more than 50 independent cDNA clones. The 5' non-coding region of the coding (sense) strand spans over 166 nucleotides, followed by one long open reading frame (ORF) of 13716 nucleotides that encodes a large putative polyprotein of 4572 amino acid residues, and by a 70-nucleotide 3' non-coding region. This ORF is apparently the longest reported to date in the plant kingdom. Amino acid sequence comparisons revealed that the large putative polyprotein includes an RNA helicase-like domain and an
RNA-dependent RNA polymerase
(replicase)-like domain. Comparisons of the amino acid sequences of these two domains and of the entire genetic organization of the rice dsRNA with those found in potyviruses and the CHV1-713 dsRNA of chestnut blight fungus suggest that the rice dsRNA is located evolutionarily between potyviruses and the CHV1-713 dsRNA. This plasmid-like dsRNA in rice seems to constitute a novel RNA replicon in plants.
Mol
Gen Genet 1995 Aug 21
PMID:Double-stranded RNA in rice: a novel RNA replicon in plants. 756 98
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