An NSP4‐dependant mechanism by which rotavirus impairs lactase enzymatic activity in brush border of human enterocyte‐like Caco‐2 cells

• Published in: Cellular microbiology Vol. 9; no. 9; pp. 2254 - 2266
• Main Authors: Beau, Isabelle, Cotte‐Laffitte, Jacqueline, Géniteau‐Legendre, Monique, Estes, Mary K., Servin, Alain L.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.09.2007; Hindawi Limited; Wiley
• Subjects: Animals; Antigens, Viral - genetics; Antigens, Viral - metabolism; Caco-2 Cells - cytology; Caco-2 Cells - enzymology; Caco-2 Cells - microbiology; Glycoproteins - genetics; Glycoproteins - metabolism; Human health and pathology; Human rotavirus; Humans; Infectious diseases; Lactase-Phlorizin Hydrolase - metabolism; Life Sciences; Macaca mulatta; Microbiology and Parasitology; Microvilli - enzymology; Microvilli - microbiology; Receptors, Virus - genetics; Receptors, Virus - metabolism; Rotavirus; Rotavirus - metabolism; Rotavirus Infections - enzymology; Toxins, Biological - genetics; Toxins, Biological - metabolism; Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - metabolism
• ISSN: 1462-5814; 1462-5822
• DOI: 10.1111/j.1462-5822.2007.00956.x

Summary Lactase‐phlorizin hydrolase (LPH, EC 3.2.1.23‐62) is a brush border membrane (BBM)‐associated enzyme in intestinal cells that hydrolyse lactose, the most important sugar in milk. Impairing in lactase activity during rotavirus infection has been described in diseased infants but the mechanism by which the functional lesion occurs remains unknown. We undertook a study to elucidate whether rotavirus impairs the lactase enzymatic activity in BBM of human enterocyte cells. In this study we use cultured human intestinal fully differentiated enterocyte‐like Caco‐2 cells to demonstrate how the lactase enzymatic activity at BBM is significantly decreased in rhesus monkey rotavirus (RRV)‐infected cells. We found that the decrease in enzyme activity is not dependent of the Ca2+‐ and cAMP‐dependent signalling events triggered by the virus. The LPH biosynthesis, stability, and expression of the protein at the BBM of infected cells were not modified. We provide evidence that in RRV‐infected cells the kinetic of lactase enzymatic activity present at the BBM was modified. Both BBMcontrol and BBMRRV have identical Km values, but hydrolyse the substrate at different rates. Thus, the BBMRRV exhibits almost a 1.5‐fold decreased Vmax than that of BBMcontrol and is therefore enzymatically less active than the latter. Our study demonstrate conclusively that the impairment of lactase enzymatic activity at the BBM of the enterocyte‐like Caco‐2 cells observed during rotavirus infection results from an inhibitory action of the secreted non‐structural rotavirus protein NSP4.