PCR cloning of a histone H1 gene from Anopheles stephensi mosquito cells: comparison of the protein sequence with histone H1-like, C-terminal extensions on mosquito ribosomal protein S6

Zhai, Yongjiao; Fallon, Ann M.

doi:10.1186/1471-2164-6-8

Cited by 5 publications

(1 citation statement)

References 19 publications

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“…Over the years, my students, postdocs and technical staff cloned and characterized mosquito ribosomal RNA and ribosomal protein genes [ 13 , 14 , 15 , 16 , 17 ] and explored aspects of their expression in the context of mosquito vitellogenesis [ 18 , 19 , 20 , 21 ] and their applications in phylogenetic analyses [ 22 ]. With cultured cells, I began to explore aspects of nucleic acid and protein metabolism [ 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 ], including characterization of genes encoding antimicrobial peptides [ 31 , 32 , 33 ]. An assigned lecture on the cell cycle and cancer informed later research with long-term associate Anna Gerenday, who used hydroxyurea to synchronize the mosquito cell cycle [ 34 ] and found a link between ecdysone and the cell cycle inhibitor protein, Dacapo [ 35 ].…”

Section: Introductionmentioning

confidence: 99%

From Mosquito Ovaries to Ecdysone; from Ecdysone to Wolbachia: One Woman’s Career in Insect Biology

Fallon

2022

Insects

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In anautogenous mosquitoes, synchronous development of terminal ovarian follicles after a blood meal provides an important model for studies on insect reproduction. Removal and implantation of ovaries, in vitro culture of dissected tissues and immunological assays for vitellogenin synthesis by the fat body showed that the Aedes aegypti (L.) (Diptera, Culicidae) mosquito ovary produces a factor essential for egg production. The discovery that the ovarian factor was the insect steroid hormone, ecdysone, provided a model for co-option of the larval hormones as reproductive hormones in adult insects. In later work on cultured mosquito cells, ecdysone was shown to arrest the cell cycle, resulting in an accumulation of diploid cells in G1, prior to initiation of DNA synthesis. Some mosquito species, such as Culex pipiens L. (Diptera, Culicidae), harbor the obligate intracellular bacterium, Wolbachia pipientis Hertig (Rickettsiales, Anaplasmataceae), in their reproductive tissues. When maintained in mosquito cell lines, Wolbachia abundance increases in ecdysone-arrested cells. This observation facilitated the recovery of high levels of Wolbachia from cultured cells for microinjection and genetic manipulation. In female Culex pipiens, it will be of interest to explore how hormonal cues that support initiation and progression of the vitellogenic cycle influence Wolbachia replication and transmission to subsequent generations via infected eggs.

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Section: Introductionmentioning

confidence: 99%

From Mosquito Ovaries to Ecdysone; from Ecdysone to Wolbachia: One Woman’s Career in Insect Biology

Fallon

2022

Insects

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The C-Terminal Extension That Characterizes Mosquito (Diptera: Culicidae) Ribosomal Protein S6 Is Widespread Among the Culicomorpha

Fallon¹,

Li²

2007

J Med Entomol

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Eukaryotic ribosomal proteins, which participate in the structure and function of the translational machinery, are generally well conserved. In Aedes and Anopheles mosquitoes (Diptera: Culicidae), however, ribosomal protein S6 (RpS6) contains a low-complexity, C-terminal extension that is absent from its homolog in Drosophila melanogaster Meigen and Manduca sexta (L.). To explore the distribution of RpS6 C-terminal extensions in genera phylogenetically closer to mosquitoes than is Drosophila, we recovered cDNAs encoding RpS6 from a phantom midge (Chaoborus sp.), a midge (Chironomus sp.), a blackfly (Simulium sp.), and a phantom crane fly [Bittacomorpha clavipes (F.) ]. Sequences of deduced translation products showed that RpS6 extensions occurred only in members of the Culicomorpha, and they were absent from B. clavipes, a member of the Ptychopteromorpha, which forms the sister group to the Culicomorpha. Likewise, the extension did not occur in Telamtoscopus sp., a moth fly classified among the more distantly related Psychodomorpha. The C-terminal extensions on RpS6 ranged in length from 81 to 190 amino acids, they were highly enriched for lysine and alanine, and they seem to be evolving more rapidly than the conventional portion of the RpS6 protein shared by all eukaryotes. Although analysis of RpS6 protein was consistent with analyses based on ribosomal DNA, suggesting that Chironomidae is the sister group to the remaining families in the Culicomorpha, trees generated from RpS6 amino acid sequences were largely congruent with accepted phylogenies based on morphological characters. Our results suggest that a C-terminal, lysinerich extension on RpS6 is a potential molecular synapomorphy for the Culicomorpha.

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Ribosomal Protein S6 Associates with Alphavirus Nonstructural Protein 2 and Mediates Expression from Alphavirus Messages

Montgomery

Berglund

Beard

et al. 2006

J Virol

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Although alphaviruses dramatically alter cellular function within hours of infection, interactions between alphaviruses and specific host cellular proteins are poorly understood. Although the alphavirus nonstructural protein 2 (nsP2) is an essential component of the viral replication complex, it also has critical auxiliary functions that determine the outcome of infection in the host. To gain a better understanding of nsP2 function, we sought to identify cellular proteins with which Venezuelan equine encephalitis virus nsP2 interacted. We demonstrate here that nsP2 associates with ribosomal protein S6 (RpS6) and that nsP2 is present in the ribosome-containing fractions of a polysome gradient, suggesting that nsP2 associates with RpS6 in the context of the whole ribosome. This result was noteworthy, since viral replicase proteins have seldom been described in direct association with components of the ribosome. The association of RpS6 with nsP2 was detected throughout the course of infection, and neither the synthesis of the viral structural proteins nor the presence of the other nonstructural proteins was required for RpS6 interaction with nsP2. nsP1 also was associated with RpS6, but other nonstructural proteins were not. RpS6 phosphorylation was dramatically diminished within hours after infection with alphaviruses. Furthermore, a reduction in the level of RpS6 protein expression led to diminished expression from alphavirus subgenomic messages, whereas no dramatic diminution in cellular translation was observed. Taken together, these data suggest that alphaviruses alter the ribosome during infection and that this alteration may contribute to differential translation of host and viral messages.The Alphavirus genus contains over 25 recognized viruses with a wide geographic distribution. Venezuelan equine encephalitis virus (VEE) is a New World alphavirus that is maintained in nature by cycling between a mosquito vector and susceptible vertebrate hosts. VEE is responsible for periodic outbreaks of disease in humans and equines and is classified as a select agent, making it a prominent pathogen among the alphaviruses.Alphaviruses possess a genome of single-stranded messagesense RNA that is approximately 11.5 kb in length. The alphavirus genome is organized such that the 5Ј two-thirds of the genome encodes four nonstructural proteins (nsP1 through nsP4), whereas the 3Ј one-third encodes three mature structural proteins (capsid, E2, and E1) that are expressed at high levels after transcription from an internal 26S subgenomic mRNA promoter. The viral genome appears to be similar to cellular mRNAs, since it contains 5Ј and 3Ј untranslated regions, with a 5Ј-terminal methylguanylate cap and a 3Ј-terminal polyadenylate tail. Upon release into a host cell, the viral genomic RNA is directly translated by the cellular translation machinery. The nonstructural proteins are synthesized as two polyproteins, termed P123 and P1234. P123 results when translation terminates at an opal termination codon between nsP3 and nsP4. When t...

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PCR cloning of a histone H1 gene from Anopheles stephensi mosquito cells: comparison of the protein sequence with histone H1-like, C-terminal extensions on mosquito ribosomal protein S6

Cited by 5 publications

References 19 publications

From Mosquito Ovaries to Ecdysone; from Ecdysone to Wolbachia: One Woman’s Career in Insect Biology

From Mosquito Ovaries to Ecdysone; from Ecdysone to Wolbachia: One Woman’s Career in Insect Biology

The C-Terminal Extension That Characterizes Mosquito (Diptera: Culicidae) Ribosomal Protein S6 Is Widespread Among the Culicomorpha

Ribosomal Protein S6 Associates with Alphavirus Nonstructural Protein 2 and Mediates Expression from Alphavirus Messages

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