Synthesis and Degradation of Ribosomes in the Mosquito This renewal application builds upon a longstanding interest in regulation of ribosome biosynthesis during mosquito reproduction. Our goals are defined in the broader context of overall efforts by many laboratories to develop molecular tools to interfere with transmission of disease by vector mosquitoes. The research has been facilitated by effective integration of in vitro studies using cultured mosquito cell lines and in vivo studies with vitellogenic mosquitoes, and includes some of the earliest applications of gene transfer technology to mosquito cells in culture. In continuing this work, we plan to focus particularly on ribosomal protein S6, the major phosphorylated protein on the eukaryotic ribosome. Based on an existing cDNA, we will use recombinant DNA techniques including the polymerase chain reaction to complete the rpS6 genomic DNA structure, including 1 to 2 kb upstream and downstream of the coding region. In this analysis we will learn whether mosquito rpS6 genes resemble the Drosophila homolog in having alternative third exons. Using primers based on the cDNA, we will undertake a limited phylogenetic analysis to explore whether a previously undescribed C-terminal similarity to histone H1 protein is shared among two closely related groups of insect vectors. The 5'-flanking sequences of the mosquito rpS6 gene will be compared to those of rpL8 and rpL34, and sites common to all three genes will be defined by functional assays in transfected cells. Proteins that interact with shared regulatory elements will be identified using gel retardation and related techniques. We will identify the sites of rpS6 phosphorylation, and investigate conditions for manipulating the state of rpS6 protein phosphorylation in cultured cells and in mosquito fat body. In addition, we will examine whether the histone-like C-terminal extension encoded by the rpS6 cDNA is phosphorylated at serine residues, and whether it is processed to generate a small, phosphorylated ribosomal protein that we have previously described in mosquito fat body. Physiological studies in the female mosquito will integrate our understanding of ribosomal protein and rRNA gene expression with the well-studied hormonal events that orchestrate the reproductive cycle. These studies provide cloned genes, molecular approaches, and characterization of regulatory processes that can potentially be manipulated to interfere with disease transmission by blood-feeding arthropods.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
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Special Emphasis Panel (ZRG1-TMP (01))
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Aultman, Kathryn S
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University of Minnesota Twin Cities
Schools of Earth Sciences/Natur
United States
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Fallon, Ann M; Gerenday, Anna (2010) Ecdysone and the cell cycle: investigations in a mosquito cell line. J Insect Physiol 56:1396-401
Fallon, Ann M; Li, Lei (2007) The C-terminal extension that characterizes mosquito (Diptera: Culicidae) ribosomal protein S6 is widespread among the Culicomorpha. J Med Entomol 44:608-16
Hernandez, Vida P; Fallon, A M (2007) Histone H1-like, lysine-rich low complexity amino acid extensions in mosquito ribosomal proteins RpL23a and RpS6 have evolved independently. Arch Insect Biochem Physiol 64:100-10
Zhai, Yongjiao; Fallon, Ann M (2005) 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. BMC Genomics 6:8
Li, Lei; Fallon, A M (2005) Recovery of cDNAs encoding ribosomal proteins S9 and L26 from Aedes albopictus mosquito cells and identification of their homologs in the malaria vector, Anopheles gambiae. Arch Insect Biochem Physiol 60:44-53
Hernandez, Vida P; Higgins, LeeAnn; Schwientek, Melinda Sue et al. (2003) The histone-like C-terminal extension in ribosomal protein S6 in Aedes and Anopheles mosquitoes is encoded within the distal portion of exon 3. Insect Biochem Mol Biol 33:901-10
Jayachandran, Gitanjali; Fallon, Ann M (2003) Ribosomal protein P0 from Aedes albopictus mosquito cells: cDNA cloning and analysis of expression. Genetica 119:1-10
Schwientek, Melinda Sue; Higgins, LeeAnn; Fallon, Ann Marie (2002) Cultured Aedes albopictus mosquito cells accumulate elongation factor-1 alpha (EF-1 alpha) during serum starvation. Insect Biochem Mol Biol 32:1055-63
Gerenday, A; Shih, K M; Herman, C C et al. (2001) Increased ribonucleotide reductase activity in hydroxyurea-resistant mosquito cells. Arch Insect Biochem Physiol 46:19-25
Niu, L L; Fallon, A M (2000) Differential regulation of ribosomal protein gene expression in Aedes aegypti mosquitoes before and after the blood meal. Insect Mol Biol 9:613-23

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