Meghwanshi, K. K. | bioRxiv Science

bioRxiv2024

TSSK homologue regulates the expression of protamine and mosquito testes specific genes in Anopheles stephensi

The testis-specific serine/threonine-protein kinase (tssk) gene is known to play an important role in spermiogenesis in mammals. This study identifies tssk homologues, As_tssk3 and Aea_tssk1 in Anopheles stephensi and Aedes aegypti, respectively. Both As_tssk3 and Aea_tssk1 were found to express in a male-specific manner throughout the development. Efficient knockdown in the expression of As_tssk3 gene was observed upon feeding An. stephensi larvae with target dsRNA. As_tssk3 knockdown led to a reduced sperm reservoir in male adults. Additionally, the knockdown of As_tssk3 resulted in the reduced expression of two male specifically expressed genes, protamine & mosquito testes specific (mts- a sperm-specific marker) homologues of An. stephensi. In-silico interaction analysis suggests that As_TSSK3 protein interacts with the Nucleosome Assembly Protein1 like-4 (NAP1like-4), a putative transcription factor. We hypothesize that As_TSSK3 regulates the transcription of protamine and mts genes via NAP, as both NAP and protamine have previously been shown to function in the establishment and maintenance of chromatin dynamics, in independent studies. The study presented here is the first report of characterisation of tssk homologues in any mosquito species. The knockout studies and phosphorylation assays would provide more insights to explore the mechanism by which As_tssk3 regulates protamine and mts genes in Anopheles stephensi. HighlightsO_LIThis study identifies the tssk homologues in Ae. aegypti and An. stephensi and showed their male-specific expression in both the mosquito species. C_LIO_LIdsRNA mediated knockdown of As_TSSK3 resulted into the males with a reduced size of sperm reservoir. C_LIO_LIdsRNA mediated knockdown of As_TSSK3 led to the reduction in the expression of As_prot and As_mts, homologues of protamine and mosquito testis specific, genes in An. stephensi. C_LIO_LIIn-silico studies suggest that As_TSSK3 regulates the expression of As_prot and As_mts genes via NAP. C_LI

genetics↗

bioRxiv2023

A sex-specific homologue of snake waprin is required for embryonic development in the Red Flour Beetle, Tribolium castaneum

1.Waprin, an extracellular secretory protein, is widely known for its antibacterial activity. Through the analysis of sex-specific transcriptomes of adult Tribolium castaneum, we identified a homologue of snake waprin (Tc_WapF) with significantly higher level of its transcripts in females. Developmental- and tissue-specific profiling revealed the widespread expression of Tc_WapF in adult female tissues. Interestingly, Tc_WapF expression is not regulated by the classical sex-determination cascade of T. castaneum, as we fail to get any attenuation in Tc_WapF transcript levels in Tcdsx and Tctra (key players of sex determination cascade of T. castaneum) knockdown females. The eggs laid by Tc_WapF knockdown females never hatched, indicating the involvement of Tc_WapF in the embryonic development in T. castaneum. This is the first report of identification a sex-specific waprin homologue in an insect and its involvement as a novel candidate in the embryonic development. Future investigations on the functional conservation of insect waprins and their mechanistic role in embryonic development can be exploited for improving pest management strategies. Summary statementTribolium castaneum possesses a female-specific homologue of snake Waprin (Tc_WapF) which is essential for its embryonic development. Expression of Tc_WapF is independent of Tribolium sex-determination cascade.

molecular biology↗