Opportunistic bacteria Serratia proteamaculans regulate the intensity of their invasion by increasing the expression of host cell surface receptors

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The Serratia proteamaculans are able to penetrate eukaryotic cells. One of the virulence factors of these bacteria is the bacterial surface protein OmpX. The OmpX protein increases the adhesion of bacteria to the surface of eukaryotic cells. In addition, this protein increases the gene expression of the EGF receptor and β1 integrin, which determine the intensity of S. proteamaculans invasion. We show that OmpX also increases the expression of E-cadherin, which is involved in S. proteamaculans invasion. The objective of this work was to compare the effect of bacteria at different growth stages on the gene expression of receptors in carcinoma cells, which normally synthesize different numbers of receptors involved in invasion. Bacteria were used after 24 hours of growth, when they had not yet synthesized the OmpX-cleaving protease protealysin, and after 48 hours of growth, when active protealysin was detected in bacterial extracts. After 24 and 48 hours of growth, the bacteria induce an increase in the gene expression of EGF receptor, E-cadherin, β1 and α5 integrins in M-HeLa cervical carcinoma cells, A549 lung carcinoma cells, Caco-2 colon adenocarcinoma cells, and DF-2 skin fibroblasts. The intensity of the increase in receptor expression depends on the properties of the cell line and the growth stage of the bacteria. Moreover, infection with S. proteamaculans causes a similar increase in the expression of only the EGF receptor and β1 integrin. Using quantitative invasion, it was shown that the intensity of bacterial invasion, depending on the growth stage of the bacterial culture, correlates with the dynamics of increased gene expression of the EGF receptor and β1 integrin. When analyzing the number of receptors, it was shown that an increase in the gene expression of the EGF receptor and β1 integrin in cells may be necessary to replenish the pool of receptors that move from the membrane into the cytoplasm of the host cell during infection. Thus, as a result of contact of the bacterial surface protein OmpX with the surface of a human cell, receptors involved in S. proteamaculans invasion accumulate. Moreover, it is the increase in the gene expression of the EGF receptor and β1-integrin that determines the sensitivity of infected cells to S. proteamaculans.

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Sobre autores

O. Tsaplina

Institute of Cytology of the Russian Academy of Sciences

Autor responsável pela correspondência
Email: olga566@mail.ru
Rússia, Saint Petersburg, 194064

Bibliografia

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2. Fig. 1. The effect of contact with OmpX-synthesizing bacteria on the expression of the E-cadherin gene by the host cell. a – in M-HeLa cells after 2-hour incubation with S. proteamaculans and Esherichia coli (OmpX) bacteria at the 48-hour growth stage, E.coli (pET21) was used as a control for bacteria that do not synthesize OmpX. b is the same, but S. proteamaculans bacteria are at the growth stages of 24 and 48 h. The control is uninfected cells. GAPDH and β-actin were used as internal controls. The difference with the expression of E-cadherin in uninfected cells was considered significant at *P < 0.05.

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3. Fig. 2. Expression of β1-integrin (a), EFP receptor (b), and α5-integrin (c) in eukaryotic cells in contact with S. proteamaculans bacteria. Bacteria were added to the cells after 24 and 48 hours of growth for 2 hours. The control was uninfected cells. GAPDH and β-actin were used as internal controls. The difference with the expression of receptors in uninfected cells was considered significant at *P < 0.05.

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4. Fig. 3. The intensity of invasion of S. proteamaculans bacteria into eukaryotic cells. Bacteria at the growth stages of 24 and 48 hours were added to eukaryotic cells for 2 hours. The invasion rate of 24-hour bacteria was assumed to be 100%. The difference in the intensity of bacterial invasion at different growth stages was considered significant at *P < 0.05.

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5. Fig. 4. Expression (a, RT-PCR) and content (b, Western blot) of a number of proteins (EGF receptor, fibronectin, β1- and α5-subunits of integrin, E-cadherin) in A549 cells upon their contact with S. proteamaculans bacteria. Bacteria at the 48 h growth stage were added to eukaryotic cells for 2 h. The control was uninfected cells. β-Actin (RT-PCR) and GAPDH (Western blotting) were used as internal controls. The difference in expression relative to uninfected cells was considered significant at *P < 0.05.

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