Molecular cloning and characterisation of SmSLK, a novel Ste20-like kinase in Schistosoma mansoni

作者：Yan Yutao; Tulasne David; Browaeys Edith; Cailliau Katia; Khayath Naji; Pierce Raymond J; Trolet Jacques; Fafeur Veronique; Ben Younes Amena; Dissous Colette^*

来源：International Journal for Parasitology, 2007, 37(14): 1539-1550.

DOI：10.1016/j.ijpara.2007.06.001

• Ste20 kinase
• Schistosoma mansoni
• mitogen-activated protein kinase
• signalling pathway

Serine/threonine kinases of the Ste20 group play important roles in various cellular functions such as growth, apoptosis and morpho-genesis. This family includes p21-Activated Kinases (PAKs) and Germinal Center Kinases (GCKs) families which contain their kinase domain in the C-terminal and N-terminal position, respectively. Here, we report the characterisation of a novel Ste20-like kinase (SLK) in the helminth parasite Schistosoma mansoni (SmSLK). SmSLK belongs to the GCK subfamily and contains a conserved N-terminal Ste20-like catalytic domain and C-terminal coiled-coil structures homologous to mammalian Lymphocyte Oriented Kinase (LOK) and SLK kinases and described as regulatory domains in these proteins. Gene assembly was performed using S. mansoni sequences available from genomic databases and indicated that SmSLK is composed of 18 exons and present in one copy in the S. mansoni genome. RT-PCR experiments demonstrated an alternative splicing of SmSLK in the exon 9 encoding the hinge region between kinase and coiled-coil domains of SmSLK and showed the expression of both transcript isoforms (SmSLK and SmSLK-S in which exon 9 is deleted) in all the S. mansoni parasite stages. Most of the Ste20-related proteins are active kinases known to regulate mitogen-activated protein kinase (MAPK) cascades. We demonstrated the kinase activity of SmSLK and SmSLK-S and their capacity to activate the MAPK/Jun N-terminal kinase (JNK) pathway in human embryonic kidney (HEK) cells as well as in Xenopus oocytes. Immunofluorescence studies indicated that SmSLK proteins were abundant in the tegument of adult schistosomes. Therefore, these results indicate that SmSLK is a new member of the GCK protein family that could participate in the regulation of MAPK cascade activation during host-parasite interactions.

• 出版日期2007-12