Molecular characterization of iron binding proteins from Glossina morsitans morsitans (Diptera: Glossinidae).

The regulation of iron is critical for maintaining homeostasis in the tsetse fly (Diptera: Glossinidae), in which both adult sexes are strict blood feeders. We have characterized the cDNAs for two putative iron-binding proteins (IBPs) involved in transport and storage; transferrin (GmmTsf1) and ferritin from Glossina morsitans morsitans. GmmTsf1 transcripts are detected in the female fat body and in adult reproductive tissues, and only in the adult developmental stage in a bloodmeal independent manner. In contrast, the ferritin heavy chain (GmmFer1HCH) and light chain (GmmFer2LCH) transcripts are expressed ubiquitously, suggesting a more general role for these proteins in iron transport and storage. Protein domain predictions for each IBP suggest both the conservation and loss of several motifs present in their vertebrate homologues. In concert with many other described insect transferrins (Tfs), putative secreted GmmTsf1 maintains 3 of the 5 residues necessary for iron-binding in the N-terminal lobe, but exhibits a loss of this iron-binding ability in the C-terminal lobe as well as a loss of large sequence blocks. Both putative GmmFer1HCH and GmmFer2LCH proteins have signal peptides, similar to other insect ferritins. GmmFer2LCH has lost the 5'UTR iron-responsive element (IRE) and, thus, translation is no longer regulated by cellular iron levels. On the other hand, GmmFer1HCH maintains both the conserved ferroxidase center and the 5'UTR IRE; however, transcript variants suggest a more extensive regulatory mechanism for this subunit.

Molecular aspects of viviparous reproductive biology of the tsetse fly (Glossina morsitans morsitans): regulation of yolk and milk gland protein synthesis.

Tsetse fly (Diptera: Glossinidae) viviparous reproductive physiology remains to be explored at the molecular level. Adult females carry their young in utero for the duration of embryonic and larval development, all the while supplying their offspring with nutrients in the form of a "milk" substance secreted from a modified accessory gland. Flies give birth to fully developed third instar larvae that pupariate shortly after birth. Here, we describe the spatial and temporal expression dynamics of two reproduction-associated genes and their products synthesized during the first and second gonotrophic cycles. The proteins studied include a putative yolk protein, Glossina morsitans morsitans yolk protein 1 (GmmYP1) and the major protein found in tsetse "milk" secretions (Glossina morsitans morsitans milk gland protein, GmmMGP). Developmental stage and tissue-specific expression of GmmYP1 show its presence exclusively in the reproductive tract of the fly during oogenesis, suggesting that GmmYP1 acts as a vitellogenic protein. Transcripts for GmmMGP are present only in the milk gland tissue and increase in coordination with the process of larvigenesis. Similarly, GmmMGP can be detected at the onset of larvigenesis in the milk gland, and is present during the full duration of pregnancy. Expression of GmmMGP is restricted to the adult stage and is not detected in the immature developmental stages. These phenomena indicate that the protein is transferred from mother to larvae as nourishment during its development. These results demonstrate that both GmmYP1 and GmmMGP are involved in tsetse reproductive biology, the former associated with the process of oogenesis and the latter with larvigenesis.