Genes in the human genome: Conservation and differentiation patterns predict a wide array of overlapping and specialized functions-1

Genes with the gene from which they originated, indicated by an asterisk. A similar tree (not shown) was obtained using the maximum-likelihood approach implemented in the program PHYML [40]. See legend for Figure 1 and text for details on Methods.<p><b>Copyright information:</b></p><p>Taken from "genes in the human genome: Conservation and differentiation patterns predict a wide array of overlapping and specialized functions"</p><p>http://www.biomedcentral.com/1471-2148/8/19</p><p>BMC Evolutionary Biology 2008;8():19-19.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2266713.</p><p></p

Genes in the human genome: Conservation and differentiation patterns predict a wide array of overlapping and specialized functions-3

(bacterial in black, and archaeal in blue) group. Human proteins are in red.<p><b>Copyright information:</b></p><p>Taken from "genes in the human genome: Conservation and differentiation patterns predict a wide array of overlapping and specialized functions"</p><p>http://www.biomedcentral.com/1471-2148/8/19</p><p>BMC Evolutionary Biology 2008;8():19-19.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2266713.</p><p></p

Genes in the human genome: Conservation and differentiation patterns predict a wide array of overlapping and specialized functions-4

D for Figure 3 and text for methods and other details. Homologs of human HSPA12A and HSPA12B were found only in vertebrates. The evolutionary tree suggests that the human genes originated from a unique progenitor also appearing in fish possibly due to a duplication event that occurred in Tetrapoda before the divergence of Amphibia. Only one HSPA12 copy was found in reptiles and birds, suggesting gene loss (or extreme divergence) of one HSPA12 copy from this lineage.<p><b>Copyright information:</b></p><p>Taken from "genes in the human genome: Conservation and differentiation patterns predict a wide array of overlapping and specialized functions"</p><p>http://www.biomedcentral.com/1471-2148/8/19</p><p>BMC Evolutionary Biology 2008;8():19-19.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2266713.</p><p></p

Genes in the human genome: Conservation and differentiation patterns predict a wide array of overlapping and specialized functions-0

Kets to the right. Alignments were obtained with Clustal W [42] and the evolutionary tree was obtained using the neighbor-joining algorithm [38] with the distance transformation method of Kimura [39], as implemented in Clustal W. A similar tree (not shown) was obtained using the maximum likelihood approach implemented in the program PHYML [40].<p><b>Copyright information:</b></p><p>Taken from "genes in the human genome: Conservation and differentiation patterns predict a wide array of overlapping and specialized functions"</p><p>http://www.biomedcentral.com/1471-2148/8/19</p><p>BMC Evolutionary Biology 2008;8():19-19.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2266713.</p><p></p

Potential effects of anti-CT-Hsp60 antibodies generated during persistent CT infections.

<p>CT-Hsp60 is released from cells infected with <i>Chlamydia trachomatis</i> (CT), and anti-CT-Hsp60 antibodies are produced by the host's immune system. In turn, these antibodies recognize surface Hsp60 on either stressed or tumor cells and, consequently, cell lysis and organ destruction can occur, determining pathogenesis of a number of diseases (see text for further details). Likewise, immunocomplexes formed by anti-CT-Hsp60 antibodies and CT- (or human-, not shown) Hsp60 can form deposits in the glomerular basal membrane, causing an idiopathic form of glomerulonephritis. Tumor cell lysis can arrest tumorigenesis, in which case it is likely that the infected individual escapes from cancer without having experienced a detectable pathology or symptom.</p

Pathologic Conditions in Which Surface Hsp60 Has Been Correlated with Pathogenesis.

<p>Pathologic Conditions in Which Surface Hsp60 Has Been Correlated with Pathogenesis.</p

Comparison between the structures of <i>Chlamydia trachomatis</i> (ct-) and human- (h-) Hsp60.

<p>Shown are the positions of the four epitopes with 100% homologies. Circle: apical domain; arrow: intermediate domain; arrowhead: equatorial domain. See text and reference <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000552#ppat.1000552-Witkin1" target="_blank">[94]</a> for further details. The images were created with PyMol (<a href="http://pymol.sourceforge.net" target="_blank">http://pymol.sourceforge.net</a>).</p

Effects of protein-secretion inhibitors on Hsp60 secretion by tumor cells.

<p><b>A</b>) Hsp60 and Hsp70 detected by Western blotting in: (<b>a</b>) immunoprecipitates from conditioned media from untreated (Unt) and inhibitor-treated H292 tumor cells; and (<b>b</b>) whole-cell lysates from H292 cells. The inhibitors are listed on top of the respective lanes. Histograms to the right represent the levels of the Hsps in immunoprecipitates determined in three separate experiments as mean percentages +/− SD of arbitrary units (AU) obtained with NIH image J 1.40 analysis software. * and ** significantly different from untreated control, p<0.005 and p<0.001, respectively. The two inhibitors (listed below the bars) significantly decreased secretion of Hsp60 and Hsp70. Also, the data from whole-cell lysates show that the protein-secretion inhibitors had no detectable effect on Hsp levels inside the cells. <b>B</b>) Hsp60 levels secreted by the H292 tumor cells before and after exposure for 1 hour, followed by a 4 hours recovery period, to protein-secretion inhibitors measured by ELISA in: (<b>a</b>) conditioned media; and (<b>b</b>) exosomes. Histograms represent Hsp60 levels expressed as pg of protein normalized for mL normalised for 10⁶ cells. Data represent mean +/− SD of three different experiments in duplicate. * Significantly different from untreated control, p<0.005. The results, which are in agreement with those obtained by Western blotting, show that the inhibitors tested significantly reduced secretion of Hsp60 by the H292 tumor cells.</p

Golgi involvement in Hsp60 secretion from tumor cells. A.

<p>TEM<b>-</b>Immunogold shows Hsp60 (black dots) in tumor cells, including in the Golgi (framed by a dashed rectangle). The arrows show the plasma membrane and arrowheads indicate Hsp60 in it. Bar: 100 nm. <b>B.</b> Extracellular levels of Hsp60 decrease after Brefeldin A (BFA) treatment of the tumor cells, as measured in immunoprecipitates from the conditioned medium (left upper panel). In contrast, Hsp70 levels are not influenced by BFA treatment (left lower panel). Right hand panel: histograms showing densitometric measurements of the Western blots to the left. UT, untreated; A.U.: arbitrary units; asterisk indicates p<0.005. <b>C.</b> ELISA results demonstrate a reduction of Hsp60 levels in the conditioned culture medium from tumor cells after BFA treatment. UT: untreated. <b>D.</b> ELISA results show a reduction of Hsp60 levels in exosomes after BFA treatment of the tumor cells. UT: untreated. Asterisks in C and D, p<0.05. Error bars represent SD.</p

Hsp60 is integrated in the membrane of exosomes from tumor cells. A–C.

<p>Illustrative data on the exosome preparations utilized in this work. <b>A</b>, Transmission electron microscopy demonstrates that the dimension of isolated vesicles is equal to, or smaller than 100 nm, which is consistent with exosomes. Bar: 100 nm. <b>B</b> and <b>C</b>, acetylcholinesterase (AChEase) and ATPse enzymatic activities, respectively, typical of isolated exosomal vesicles, compared to control (conditioned culture medium). In B, the solid line represents data from exosomes, and the broken line represents results from conditioned culture medium. Vertical axis, AChEase activity in arbitrary units (AU), reflecting 412 nm absorbance; horizontal axis, time of reaction in minutes. In C, 1, marker (positive control); 2, conditioned culture medium; 3, exosomes. <b>D.</b> Treatment with sodium carbonate alone (lane 3, asterisk), or in association with proteinase K buffer (lane 4), does not remove Hsp60 from the exosomes, whereas treatment with Proteinase K does (lane 2). Lane 1, untreated exosomes (positive control).</p