Molecular Networking Reveals Two Distinct Chemotypes in Pyrroloiminoquinone-Producing Tsitsikamma favus Sponges
- Kalinski, Jarmo-Charles J, Waterworth, Samantha C, Noundou, Xavier S, Jiwaji, Meesbah, Parker-Nance, Shirley, Krause, Rui W M, McPhail, Kerry L, Dorrington, Rosemary A
- Authors: Kalinski, Jarmo-Charles J , Waterworth, Samantha C , Noundou, Xavier S , Jiwaji, Meesbah , Parker-Nance, Shirley , Krause, Rui W M , McPhail, Kerry L , Dorrington, Rosemary A
- Date: 2019
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/131618 , vital:36673 , https://doi.org/10.3390/md17010060
- Description: The temperate marine sponge, Tsitsikamma favus, produces pyrroloiminoquinone alkaloids with potential as anticancer drug leads. We profiled the secondary metabolite reservoir of T. favus sponges using HR-ESI-LC-MS/MS-based molecular networking analysis followed by preparative purification efforts to map the diversity of new and known pyrroloiminoquinones and related compounds in extracts of seven specimens. Molecular taxonomic identification confirmed all sponges as T. favus and five specimens (chemotype I) were found to produce mainly discorhabdins and tsitsikammamines. Remarkably, however, two specimens (chemotype II) exhibited distinct morphological and chemical characteristics: the absence of discorhabdins, only trace levels of tsitsikammamines and, instead, an abundance of unbranched and halogenated makaluvamines. Targeted chromatographic isolation provided the new makaluvamine Q, the known makaluvamines A and I, tsitsikammamine B, 14-bromo-7,8-dehydro-3-dihydro-discorhabdin C, and the related pyrrolo-ortho-quinones makaluvamine O and makaluvone. Purified compounds displayed different activity profiles in assays for topoisomerase I inhibition, DNA intercalation and antimetabolic activity against human cell lines. This is the first report of makaluvamines from a Tsitsikamma sponge species, and the first description of distinct chemotypes within a species of the Latrunculiidae family. This study sheds new light on the putative pyrroloiminoquinone biosynthetic pathway of latrunculid sponges
- Full Text:
- Date Issued: 2019
- Authors: Kalinski, Jarmo-Charles J , Waterworth, Samantha C , Noundou, Xavier S , Jiwaji, Meesbah , Parker-Nance, Shirley , Krause, Rui W M , McPhail, Kerry L , Dorrington, Rosemary A
- Date: 2019
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/131618 , vital:36673 , https://doi.org/10.3390/md17010060
- Description: The temperate marine sponge, Tsitsikamma favus, produces pyrroloiminoquinone alkaloids with potential as anticancer drug leads. We profiled the secondary metabolite reservoir of T. favus sponges using HR-ESI-LC-MS/MS-based molecular networking analysis followed by preparative purification efforts to map the diversity of new and known pyrroloiminoquinones and related compounds in extracts of seven specimens. Molecular taxonomic identification confirmed all sponges as T. favus and five specimens (chemotype I) were found to produce mainly discorhabdins and tsitsikammamines. Remarkably, however, two specimens (chemotype II) exhibited distinct morphological and chemical characteristics: the absence of discorhabdins, only trace levels of tsitsikammamines and, instead, an abundance of unbranched and halogenated makaluvamines. Targeted chromatographic isolation provided the new makaluvamine Q, the known makaluvamines A and I, tsitsikammamine B, 14-bromo-7,8-dehydro-3-dihydro-discorhabdin C, and the related pyrrolo-ortho-quinones makaluvamine O and makaluvone. Purified compounds displayed different activity profiles in assays for topoisomerase I inhibition, DNA intercalation and antimetabolic activity against human cell lines. This is the first report of makaluvamines from a Tsitsikamma sponge species, and the first description of distinct chemotypes within a species of the Latrunculiidae family. This study sheds new light on the putative pyrroloiminoquinone biosynthetic pathway of latrunculid sponges
- Full Text:
- Date Issued: 2019
Binding and entry of a non-enveloped T=4 insect RNA virus is triggered by alkaline pH
- Penkler, David L, Jiwaji, Meesbah, Domitrovic, Tatiana, Short, James R, Johnson, John E, Dorrington, Rosemary A
- Authors: Penkler, David L , Jiwaji, Meesbah , Domitrovic, Tatiana , Short, James R , Johnson, John E , Dorrington, Rosemary A
- Date: 2016
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/65995 , vital:28875 , https://doi.org/10.1016/j.virol.2016.08.028
- Description: publisher version , Tetraviruses are small, non-enveloped, RNA viruses that exclusively infect lepidopteran insects. Their particles comprise 240 copies of a single capsid protein precursor (CP), which undergoes autoproteolytic cleavage during maturation. The molecular mechanisms of capsid assembly and maturation are well understood, but little is known about the viral infectious lifecycle due to a lack of tissue culture cell lines that are susceptible to tetravirus infection. We show here that binding and entry of the alphatetravirus, Helicoverpa armigera stunt virus (HaSV), is triggered by alkaline pH. At pH 9.0, wild-type HaSV virus particles undergo conformational changes that induce membrane-lytic activity and binding to Spodoptera frugiperda Sf9 cells. Binding is followed by entry and infection, with virus replication complexes detected by immunofluorescence microscopy within 2 h post-infection and the CP after 12 h. HaSV particles produced in S. frugiperda Sf9 cells are infectious. Helicoverpa armigera larval virus biofeed assays showed that pre-treatment with the V-ATPase inhibitor, Bafilomycin A1, resulted in a 50% decrease in larval mortality and stunting, while incubation of virus particles at pH 9.0 prior to infection restored infectivity. Together, these data show that HaSV, and likely other tetraviruses, requires the alkaline environment of the lepidopteran larval midgut for binding and entry into host cells.
- Full Text: false
- Date Issued: 2016
- Authors: Penkler, David L , Jiwaji, Meesbah , Domitrovic, Tatiana , Short, James R , Johnson, John E , Dorrington, Rosemary A
- Date: 2016
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/65995 , vital:28875 , https://doi.org/10.1016/j.virol.2016.08.028
- Description: publisher version , Tetraviruses are small, non-enveloped, RNA viruses that exclusively infect lepidopteran insects. Their particles comprise 240 copies of a single capsid protein precursor (CP), which undergoes autoproteolytic cleavage during maturation. The molecular mechanisms of capsid assembly and maturation are well understood, but little is known about the viral infectious lifecycle due to a lack of tissue culture cell lines that are susceptible to tetravirus infection. We show here that binding and entry of the alphatetravirus, Helicoverpa armigera stunt virus (HaSV), is triggered by alkaline pH. At pH 9.0, wild-type HaSV virus particles undergo conformational changes that induce membrane-lytic activity and binding to Spodoptera frugiperda Sf9 cells. Binding is followed by entry and infection, with virus replication complexes detected by immunofluorescence microscopy within 2 h post-infection and the CP after 12 h. HaSV particles produced in S. frugiperda Sf9 cells are infectious. Helicoverpa armigera larval virus biofeed assays showed that pre-treatment with the V-ATPase inhibitor, Bafilomycin A1, resulted in a 50% decrease in larval mortality and stunting, while incubation of virus particles at pH 9.0 prior to infection restored infectivity. Together, these data show that HaSV, and likely other tetraviruses, requires the alkaline environment of the lepidopteran larval midgut for binding and entry into host cells.
- Full Text: false
- Date Issued: 2016
Expanding the host range of small insect RNA viruses: Providence virus (Carmotetraviridae) infects and replicates in a human tissue culture cell line
- Jiwaji, Meesbah, Short, James R, Dorrington, Rosemary A
- Authors: Jiwaji, Meesbah , Short, James R , Dorrington, Rosemary A
- Date: 2016
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/65979 , vital:28874 , https://doi.org/10.1099/jgv.0.000578
- Description: publisher version , Tetraviruses are small, positive (+ve)-sense ssRNA viruses that infect the midgut cells of lepidopteran larvae. Providence virus(PrV) is the only member of the family Carmotetraviridae (previously Tetraviridae). PrV particles exhibit the characteristic tetraviral T=4 icosahedral symmetry, but PrV is distinct from other tetraviruses with respect to genome organization and viral non-structural proteins. Currently, PrV is the only tetravirus known to infect and replicate in lepidopteran cell culture lines. In this report we demonstrate, using immunofluorescence microscopy, that PrV infects and replicates in a human tissue culture cell line (HeLa), producing infectious virus particles. We also provide evidence for PrV replication in vitro in insect, mammalian and plant cell-free systems. This study challenges the long-held view that tetraviruses have a narrow host range confined to one or a few lepidopteran species and highlights the need to consider the potential for apparently non-infectious viruses to be transferred to new hosts in the laboratory.
- Full Text: false
- Date Issued: 2016
- Authors: Jiwaji, Meesbah , Short, James R , Dorrington, Rosemary A
- Date: 2016
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/65979 , vital:28874 , https://doi.org/10.1099/jgv.0.000578
- Description: publisher version , Tetraviruses are small, positive (+ve)-sense ssRNA viruses that infect the midgut cells of lepidopteran larvae. Providence virus(PrV) is the only member of the family Carmotetraviridae (previously Tetraviridae). PrV particles exhibit the characteristic tetraviral T=4 icosahedral symmetry, but PrV is distinct from other tetraviruses with respect to genome organization and viral non-structural proteins. Currently, PrV is the only tetravirus known to infect and replicate in lepidopteran cell culture lines. In this report we demonstrate, using immunofluorescence microscopy, that PrV infects and replicates in a human tissue culture cell line (HeLa), producing infectious virus particles. We also provide evidence for PrV replication in vitro in insect, mammalian and plant cell-free systems. This study challenges the long-held view that tetraviruses have a narrow host range confined to one or a few lepidopteran species and highlights the need to consider the potential for apparently non-infectious viruses to be transferred to new hosts in the laboratory.
- Full Text: false
- Date Issued: 2016
A broad host range reporter plasmid for the analysis of divergent promoter regions
- Jiwaji, Meesbah, Matcher, Gwynneth F, Dorrington, Rosemary A
- Authors: Jiwaji, Meesbah , Matcher, Gwynneth F , Dorrington, Rosemary A
- Date: 2008
- Language: English
- Type: Article
- Identifier: vital:6476 , http://hdl.handle.net/10962/d1006164 , http://www.scielo.org.za/scielo.php?pid=S0038-23532008000400013&script=sci_arttext
- Description: Although many vectors exist for Escherichia coli and closely related species, there are few broad host range vectors that can be conjugated into a large variety of Gram-negative bacteria. We have constructed a broad host range vector, pMJ445, that facilitates the analysis of divergent promoters in Gram-negative bacteria. The vector was validated using two intergenic regions derived from gene clusters involved in hydantoin hydrolysis, from the environmental isolates Pseudomonas putida and Agrobacterium tumefaciens. The DNA sequences analysed were capable of activating expression of the reporter enzymes, β-glucuronidase and β-galactosidase, present on pMJ445, indicating the presence of divergent promoters in the sequences selected. In addition, we demonstrated that pMJ445 can be applied to gene regulation studies.
- Full Text:
- Date Issued: 2008
- Authors: Jiwaji, Meesbah , Matcher, Gwynneth F , Dorrington, Rosemary A
- Date: 2008
- Language: English
- Type: Article
- Identifier: vital:6476 , http://hdl.handle.net/10962/d1006164 , http://www.scielo.org.za/scielo.php?pid=S0038-23532008000400013&script=sci_arttext
- Description: Although many vectors exist for Escherichia coli and closely related species, there are few broad host range vectors that can be conjugated into a large variety of Gram-negative bacteria. We have constructed a broad host range vector, pMJ445, that facilitates the analysis of divergent promoters in Gram-negative bacteria. The vector was validated using two intergenic regions derived from gene clusters involved in hydantoin hydrolysis, from the environmental isolates Pseudomonas putida and Agrobacterium tumefaciens. The DNA sequences analysed were capable of activating expression of the reporter enzymes, β-glucuronidase and β-galactosidase, present on pMJ445, indicating the presence of divergent promoters in the sequences selected. In addition, we demonstrated that pMJ445 can be applied to gene regulation studies.
- Full Text:
- Date Issued: 2008
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