Insights into the insect salivary gland proteome: diet-associated changes in caterpillar labial salivary proteins.

The primary function of salivary glands is fluid and protein secretion during feeding. Compared to mammalian systems, little is known about salivary protein secretion processes and the effect of diet on the salivary proteome in insect models. Therefore, the effect of diet nutritional quality on caterpillar labial salivary gland proteins was investigated using an unbiased global proteomic approach by nanoLC/ESI/tandem MS. Caterpillars of the beet armyworm, Spodoptera exigua Hübner, were fed one of three diets: an artificial diet containing their self-selected protein to carbohydrate (p:c) ratio (22p:20c), an artificial diet containing a higher nutritional content but the same p:c ratio (33p:30c) or the plant Medicago truncatula Gaertn. As expected, most identified proteins were associated with secretory processes and not influenced by diet. However, some diet-specific differences were observed. Nutrient stress-associated proteins, such as peptidyl-propyl cis-trans isomerase and glucose-regulated protein94/endoplasmin, and glyceraldehyde 3-phosphate dehydrogenase were identified in the labial salivary glands of caterpillars fed nutritionally poor diets, suggesting a link between nutritional status and vesicular exocytosis. Heat shock proteins and proteins involved in endoplasmic reticulum-associated protein degradation were also abundant in the labial salivary glands of these caterpillars. In comparison, proteins associated with development, such as arylphorin, were found in labial salivary glands of caterpillars fed 33p:30c. These results suggest that caterpillars fed balanced or nutritionally-poor diets have accelerated secretion pathways compared to those fed a protein-rich diet.

The p150 subunit of the histone chaperone Caf-1 interacts with the transcriptional repressor Gfi1.

Modification of histones is critically involved in regulating chromatin structure and gene expression. The zinc finger protein Gfi1 silences transcription by recruiting a complex of histone modifying enzymes such as LSD-1/CoRest and HDAC-1 to target gene promoters. Here we present evidence that Gfi1 forms a complex with the p150 subunit of the histone chaperone chromatin assembly factor-1 (Caf-1). Gfi1 and p150 interact at endogenous expression levels and co-localize in distinct sub-nuclear structures. We show that p150 enhances Gfi1-mediated transcriptional repression and that it occupies Gfi1 target gene promoters in transfected cells and primary murine T cells only in the presence of Gfi1. Finally, size exclusion chromatography shows a fraction of p150 to coelute with Gfi1, LSD-1 and HDAC-1 and thus provides evidence that p150 is part of the Gfi1 repression complex. Since p150 binds directly to histones H3 and H4, our findings suggest that p150 may link the DNA-bound Gfi1 repressor complex to histones enabling modifications required for transcriptional silencing.

Diet-specific salivary gene expression and glucose oxidase activity in Spodoptera exigua (Lepidoptera: Noctuidae) larvae.

Saliva secreted during caterpillar feeding contains enzymes to initiate digestion or detoxify noxious plant compounds. Activity of some salivary enzymes is diet-dependent and may be transcriptionally regulated. In this study, cDNA-amplified fragment length polymorphism was used to identify beet armyworm, Spodoptera exigua Hübner, labial salivary genes that are differentially expressed in response to diet. In addition, SeGOX was sequenced based on homology and characterized to confirm that the transcript encodes a functional enzyme. Three labial salivary transcripts, encoding glucose oxidase (GOX) and two proteins of unknown function (Se1H and Se2J), were expressed in a diet-specific manner. Since diet, particularly the protein to digestible carbohydrate levels and ratio, may affect labial salivary enzyme activity, the influence of nutritional quality on gene expression was determined. Transcript levels of the labial salivary genes Se1H, Se2J and SeGOX increased with dietary carbohydrate levels, regardless of protein concentrations. In contrast GOX enzymatic activity increased with increasing dietary carbohydrates when caterpillars were fed protein-rich diets, but not when caterpillars were fed protein-poor diets. Our results suggest that dietary carbohydrates affect SeGOX, Se1H and Se2J transcription, but dietary protein or amino acid levels affect translational and/or post-translational regulation of the enzyme GOX.

A variant allele of Growth Factor Independence 1 (GFI1) is associated with acute myeloid leukemia.

The GFI1 gene encodes a transcriptional repressor, which regulates myeloid differentiation. In the mouse, Gfi1 deficiency causes neutropenia and an accumulation of granulomonocytic precursor cells that is reminiscent of a myelodysplastic syndrome. We report here that a variant allele of GFI1 (GFI1(36N)) is associated with acute myeloid leukemia (AML) in white subjects with an odds ratio of 1.6 (P < 8 x 10(-5)). The GFI1(36N) variant occurred in 1806 AML patients with an allele frequency of 0.055 compared with 0.035 in 1691 healthy control patients in 2 independent cohorts. We observed that both GFI1 variants maintain the same activity as transcriptional repressors but differ in their regulation by the AML1/ETO (RUNX1/RUNX1T1) fusion protein produced in AML patients with a t(8;21) translocation. AML1/ETO interacts and colocalizes with the more common GFI1(36S) form in the nucleus and inhibits its repressor activity. However, the variant GFI1(36N) protein has a different subnuclear localization than GFI1(36S). As a consequence, AML1/ETO does not colocalize with GFI1(36N) and is unable to inhibit its repressor activity. We conclude that both variants of GFI1 differ in their ability to be regulated by interacting proteins and that the GFI1(36N) variant form exhibits distinct biochemical features that may confer a predisposition to AML.