Tripartite Motif-containing 28 Research Articles

Tripartite Motif-Containing 29 Expression in Squamous Cell Carcinoma

Dear Editor: Tripartite motif-containing 29 (TRIM29), also known as the ataxia telangiectasia group D complementing gene, is reduced in certain cancers by some reports, although, others report higher expression of TRIM29 in several cancers. While the exact functions of TRIM proteins have yet to be specified, they undergo viral replication, and participate in signal transduction and development in many human diseases, especially cancer. A few members of the TRIM family are associated with cellular growth and processes, such as transcriptional regulation, cell growth, apoptosis, development and oncogenesis1. Several TRIM proteins play many roles in the cellular processes, including the development and growth of diseases, but the physiological and pathological functions of TRIM29 have not been confirmed; while its presence in the cell membrane has been observed2. TRIM29 is typically expressed in the placenta, lung, thymus, prostate, testis, and colon, but not in the heart, brain, skeletal muscle, pancreas, spleen, ovary, or the small intestine, as measured by Northern blot analysis3. There have not been many studies on TRIM29, but it is assessed that TRIM29 expression is suppressed in breast and prostate cancer4-6. In a study by Schlomm et al.7, cDNA microarray analysis on prostatic cancer cells and noncancerous prostatic cells conducted using laser microdissection, resulted in 216 different genes, with TRIM29 as one of the downregulated genes. Similarly, Ernst et al.4 reported that TRIM29 was shown to be downregulated. Liu et al.6 reported that TRIM29 functions as a tumor suppressor in nontumorigenic breast cells and invasive estrogen receptor positive breast cancer using cDNA microarray analysis. TRIM29's functions may differ depending on the cell type, but there have not been any studies on the role of TRIM29 on tumorigenesis. TRIM29 over-expresses in lung, bladder, colorectal, ovarian, gastric, pancreatic and endometrial cancers and in multiple myeloma, and recent studies show that TRIM29 is associated with tumor invasion and lymph node metastasis. A recent report correlated TRIM29 expression in gastric cancer and poor histological grade, large tumor size, extent of tumor invasion, and lymph node metastasis8. The Institutional Review Board of Seoul Soonchunhyang University Hospital reviewed and approved this research protocol, which involved the use of tissue samples. We analyzed a total of 6 normal skin tissues and 18 malignant skin tumor tissues, including 6 malignant melanoma (MM), 6 squamous cell carcinoma (SCC), and 6 basal cell carcinoma (BCC), using Western blot and immunohistochemistry. The human MM cell line G361 served as a positive control for the TRIM29 expression. The human MM cell line G361 was not expressed for TRIM29 protein. In this study, TRIM29, which indicates the metastatic potentiality in gastric cancer, was measured in BCC, SCC and MM cancers, using Western blot and immunohistologic method. The results show that the expression of TRIM29 was not expressed in 6 cases of BCC with slow growth and poor metastasis. In 6 cases of SCC, which is aggressive and locally invasive, but not metastatic cancer, the expression of TRIM29 was strongly positive (Fig. 1A). In the 6 cases of MM, which is invasive and has distant sites metastatic potential through hematogenous and lymphagenous paths, TRIM29 protein was not expressed. Fig. 1 (A) TRIM29 protein was expressed on squamous cell carcinomal, normal skin tissue by western blot analysis. (B) The score of relative TRIM29 expression amount in western blotting. TRIM29: tripartite motif-containing 29, MM: malignant melanoma, SCC: squamous ... In all six MM and BCC, TRIM29 was not expressed (data not shown). The amount of expression in Western blotting is displayed graphically (Fig. 1B). Immunohistochemical study showed that the staining pattern of TRIM29 in normal human skin tissues mirrored those of a Western blot analysis (Fig. 2). Fig. 2 Immunohistochemical analysis of tripartite motif-containing 29 (TRIM29) expression in (A) malignant melanoma, (B) squamous cell carcinoma, (C) basal cell carcinoma, and (D) normal skin tissue. Only squamous cell carcinoma displayed TRIM29 expression. ... The exact function and mechanism of TRIM29 during tumor genesis have not been confirmed. The possible roles and mechanisms of TRIM29 protein expression in malignant skin cancers will be examined in additional studies.

TRIM29 Functions as a Tumor Suppressor in Nontumorigenic Breast Cells and Invasive ER+ Breast Cancer

Tripartite motif-containing 29 (TRIM29) is a member of the TRIM protein family that has been implicated in hematologic and solid tumor cancers. We found that TRIM29 functions as a tumor suppressor in both the nontumorigenic MCF10A [estrogen receptor (ER)-/TRIM29+] breast cell line and the invasive MCF7 (ER+/TRIM29-) breast cell line. Silencing TRIM29 in MCF10A cells resulted in preneoplastic changes that included loss of polarity in three-dimensional culture, increased proliferation, anchorage-independent growth, and increased migration and invasion. Conversely, the introduction of TRIM29 into MCF7 cells caused reversion to a less aggressive phenotype by antagonizing the growth effect of 17β-estradiol. The interaction between TRIM29 and ER signaling in MCF7 cells was supported by a reduction in ERE binding in the presence of TRIM29 and suppression of ER-dependent gene expression of TFF1, FOS, and GREB1. By microarray analyses, we showed that younger women (<55 years of age) with early-stage, ER+ breast cancer who were given no adjuvant systemic therapy had a significantly lower risk of relapse when their tumor had high TRIM29 expression (P = 0.02). This effect was not observed in older women (>55 years of age) and thus may be due to menopause and loss of circulating estrogens. Our results suggest that loss of TRIM29 expression in normal breast luminal cells can contribute to malignant transformation and lead to progression of ER+ breast cancer in premenopausal women.

Structural Basis for the Activation of Innate Immune Pattern-Recognition Receptor RIG-I by Viral RNA

RIG-I is a key innate immune pattern-recognition receptor that triggers interferon expression upon detection of intracellular 5'triphosphate double-stranded RNA (5'ppp-dsRNA) of viral origin. RIG-I comprises N-terminal caspase activation and recruitment domains (CARDs), a DECH helicase, and a C-terminal domain (CTD). We present crystal structures of the ligand-free, autorepressed, and RNA-bound, activated states of RIG-I. Inactive RIG-I has an open conformation with the CARDs sequestered by a helical domain inserted between the two helicase moieties. ATP and dsRNA binding induce a major rearrangement to a closed conformation in which the helicase and CTD bind the blunt end 5'ppp-dsRNA with perfect complementarity but incompatibly with continued CARD binding. We propose that after initial binding of 5'ppp-dsRNA to the flexibly linked CTD, co-operative tight binding of ATP and RNA to the helicase domain liberates the CARDs for downstream signaling. These findings significantly advance our molecular understanding of the activation of innate immune signaling helicases.

Identification of tripartite motif-containing 22 (TRIM22) as a novel NF-κB activator

Increasing evidence suggests that TRIM family proteins may play important roles in the regulation of innate immune signaling pathways. Here we report TRIM22 is involved in the activation of NF-κB. It was found that overexpression of TRIM22 could dose-dependently activate NF-κB as demonstrated by reporter gene assay and electrophoretic mobility shift assay, but had no effect on the activity of other transcription factors, including NF-AT, AP-1, C/EBP and IRFs. Further study showed that both the N-terminal RING domain and C-terminal SPRY domain were crucial for TRIM22-mediated NF-κB activation. Moreover, our results revealed that TRIM22 overexpression could significantly induce the secretion of pro-inflammatory cytokines by human macrophage cell line U937 in an NF-κB-dependent manner. These data suggested that TRIM22 was a positive regulator of NF-κB-mediated transcription.

TRIM24 links a non-canonical histone signature to breast cancer

Recognition of modified histone species by distinct structural domains within “reader” proteins plays a critical role in the regulation of gene expression. Readers that simultaneously recognize histones with multiple marks allow transduction of complex chromatin modification patterns into specific biological outcomes. Here, we report that chromatin regulator TRIM24 functions as a reader of dual histone marks via tandem Plant Homeodomain (PHD) and Bromodomain (Bromo). The three-dimensional structure of TRIM24 PHD-Bromo revealed a single functional unit for combinatorial recognition of unmodified H3K4 (H3K4me0) and acetylated H3K23 (H3K23ac) within the same histone tail. TRIM24 binds chromatin and estrogen receptor to activate estrogen-dependent genes associated with cellular proliferation and tumor development. Aberrant expression of TRIM24 negatively correlates with survival of breast cancer patients. The PHD-Bromo of TRIM24 provides a structural rationale for chromatin activation via a noncanonical histone signature, establishing a new paradigm by which chromatin readers may influence cancer pathogenesis.

385 A first in human phase 1 study of the safety and pharmacokinetics of a novel Cdc7 inhibitor NMS-1116354, administered orally to patients with solid tumors

Tripartite Motif Containing 11 (TRIM11), a member of TRIM proteins is overexpressed in gliomas and lung cancer. However, the role of TRIM11 in hepatocellular carcinoma (HCC) is unknown.Herein, we aimed to investigate the expression and clinical significance role of TRIM11 in HCC.In this study, our data showed significant higher TRIM11 in HCC tissues (n = 117) than in the matched non-tumor liver (NTL) tissues (P < 0.01). In consistent with above data, we also found TRIM11 protein expression was significantly increased compared with the matched NTL (P < 0.01) by immunohistochemistry analysis. Additionally, our results showed that TRIM11 protein expression in HCC tissues was significantly associated with pathological grade (P < 0.01), tumor postoperative metastasis (P = 0.031), recurrence (P = 0.022), and serum a-fetoprotein (AFP) (P < 0.01). Moreover, patients’ survival was negatively correlated with TRIM11 protein expression. Furthermore, we found that TRIM11 protein was an independent prognostic factor for disease-free (P < 0.01) and overall survival (P < 0.01) in HCC patients.Our data showed that TRIM11 expression was significantly elevated in HCC tissues. The overexpression of TRIM11 is closely associated with HCC progression and poor survival of the patients, indicating TRIM11 is a potential therapeutic target for HCC patients.

Abstract 2960: TRIM29 is a novel biomarker for basal-like breast cancer

Abstract Basal-like breast cancer is a subtype of tumors associated with aggressive clinical behavior and poor prognosis. The genes responsible for their aggressive phenotype have not been well characterized and may provide an opportunity for targeted therapies. Tripartite motif-containing 29 (TRIM29) is a member of the TRIM protein family. Microarray and RT-qPCR analyses shows TRIM29 overexpressed in Basal-like breast tumors and it is highly correlated to other basal tumor markers such as CK5/6, CK17, EGFR, and FOXC1. We recently observed using immunohistochemistry that TRIM29 predominantly localizes to the nucleus in basal tumor epithelial cells, in contrast to a cytoplasmic localization seen in ER-positive tumors. Thus, TRIM29 may act as a transcriptional regulatory factor involved in carcinogenesis. In order to investigate the role of TRIM29 in the initiation and progression of Basal-like breast cancer, we employed chromatin immunoprecipitation and massively parallel sequencing (ChIP-seq) to create a genome-wide profile of in vivo TRIM29-binding sites in MDA-MB-468 cells, a human breast cancer cell line. ChIP-seq analysis revealed 94 putative TRIM29 binding regions in the cancer genome of MDA-MB-468 cells with an estimated false discovery rate of less than 5%. One TRIM29 binding motif was found in the TRIM29 binding regions with a core consensus sequence of CTGTCTCC. We also identified some known cancer genes in close proximity to TRIM29 binding sites, including JUND, FBXO31, CCND3, EGFR, and FOXC1. In addition to mapping TRIM29-DNA binding sites, we used co-immunoprecipitation (co-IP) coupled with tandem mass spectrometry to identify proteins that interact with TRIM29 in MDA-MB-468 cells. Proteins that co-precipitated with TRIM29 included splicing factor proline/glutamine-rich (SFPQ, a transcriptional co-repressor), poly(A) binding protein (PABPC4); FUS and TAF15 RNA polymerase II (both are proto-oncoproteins and transcriptional activators), and A kinase anchor protein 13 (AKAP13, an adaptor protein involved in protein kinase A signaling pathway). Confirmation of the initial co-IP results was achieved through several independent techniques, including detection of binding partners in TRIM29 precipitates by Western blot and reciprocal co-IP using antibodies to partner proteins and detection of TRIM29 in the precipitates. Overall, results from our studies indicated that besides functioning as a transcription factor, TRIM29 may also participate in signal transduction pathways by forming protein complexes. Our studies have identified new biomarkers for basal-like breast cancers and revealed kinases that could potentially be targeted by biologic therapies. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2960.

Abstract 2372: The metastasis efficiency modifier ribosomal RNA processing 1 homologue (RRP1B) is a chromatin-associated factor

Abstract Previous experiments using the polyoma middle T (PyMT) mouse model of mammary tumorigenesis have demonstrated that host germline variation modulates metastasis in breast cancer. Recent work using the PyMT model has identified novel metastasis susceptibility genes, an example of which includes Rrp1b (ribosomal RNA processing 1 homolog B). Over-expression of Rrp1b in a mouse mammary tumor cell line induces a gene expression signature that predicts survival in a well characterized Dutch breast cancer dataset. Additionally, a polymorphism was identified within human RRP1B (G1421A; P436L), which is associated with differential survival in multiple breast cancer cohorts. Here we extend the analysis of RRP1B function by demonstrating that the Rrp1b activation gene expression signature accurately predicts survival in three additional publicly available breast carcinoma gene expression datasets. We have also investigated the mechanism of action of RRP1B by defining protein binding partners for this germline-encoded metastasis modifier. Tandem affinity purification demonstrated that RRP1B physically interacts with many nucleosome binding factors, all of which have been implicated in regulation of gene expression. Examples include the linker histone H1X, poly(ADP-ribose) polymerase 1, tripartite motif-containing 28 (TIF-1β/KAP1), and cold shock domain protein A. These interactions were confirmed by co-immunofluorescence (Co-IF) and co-immunoprecipitation (Co-IP). Additionally, co-IF experiments demonstrated that RRP1B is localized to the perinuclear and perinucleolar regions, which is a particularly interesting given that factors localized to these regions are often involved in heterochromatinization and gene silencing. Furthermore, co-IP and co-IF demonstrated that RRP1B interacts with heterochromatin protein-1α and acetyl-histone H4 lysine 5, suggesting that RRP1B may well regulate gene expression through interactions with heterochromatin and euchromatin. Finally, we investigated the effects of ectopic expression of the 436Pro and 436Leu RRP1B allelic variants that have previously been associated with differential survival in breast cancer. Gene expression analyses demonstrate that, compared with ectopic expression of wildtype 436 Pro RRP1B in HeLa cells, the variant 436Leu RRP1B differentially modulates various transcription factors controlled by TRIM28 and CSDA. These data suggest that RRP1B, a tumor progression and metastasis susceptibility candidate gene, is potentially a dynamic modulator of transcription and chromatin structure. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2372.

Gene Expression Profiles of Uterine Normal Myometrium and Leiomyoma and Their Estrogen ResponsivenessIn Vitro

Background : Uterine leiomyomas are common benign smooth muscle tumors among the reproductive aged-women. The research has been aimed to identify the differentially expressed genes between normal myometrium and leiomyoma and to investigate the effects of E 2 on their expression. Methods : Gene microarray analysis was performed to identify the differentially expressed genes between normal myomerium and leiomyoma. The data was confirmed at protein level by tissue microarray. Results : Gene microarray analysis revealed 792 upregulated genes in leiomyoma. Four genes (tropomyosin 4 [TPM4 ], collagen, type IV, alpha 2 [COL4a2], insulin-like growth factor binding protein 5 [IGFBP5 ], tripartite motif-containing 28 [TRIM28]) showed the most dramatic upregulation in all leiomyoma samples. Tissue microarray analyses of 262 sample pairs showed significantly elevated expression of TPM4, IGFBP5, estrogen receptor-a, and progesterone receptor (PR) protein in leiomyoma from the patients in their forties, COL4a2 in the forties and fifties age-groups, and TRIM28 in the thirties age-group. PR, insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R) and IGFBP5 were induced by E 2 in in vitro culture of tissue explants from which cells migrated throughout the plate. Among these, PR, IGF-1, IGFBP5 genes showed higher expression in tissue compared to cells-derived from tissue in leiomyoma and IGF-1R in leiomyoma cell. Conclusions : This observation implies the importance of the whole tissue context including the cells-derived from tissue in the research for the understanding of molecular mechanism of leiomyoma. Here, we report higher expression of TRIM28 in leiomyoma for the first time and identify E 2-responsive genes that may have important roles in leiomyoma development.

The Metastasis Efficiency Modifier Ribosomal RNA Processing 1 Homolog B (RRP1B) Is a Chromatin-associated Factor

There is accumulating evidence for a role of germ line variation in breast cancer metastasis. We have recently identified a novel metastasis susceptibility gene, Rrp1b (ribosomal RNA processing 1 homolog B). Overexpression of Rrp1b in a mouse mammary tumor cell line induces a gene expression signature that predicts survival in breast cancer. Here we extend the analysis of RRP1B function by demonstrating that the Rrp1b activation gene expression signature accurately predicted the outcome in three of four publicly available breast carcinoma gene expression data sets. In addition, we provide insights into the mechanism of RRP1B. Tandem affinity purification demonstrated that RRP1B physically interacts with many nucleosome binding factors, including histone H1X, poly(ADP-ribose) polymerase 1, TRIM28 (tripartite motif-containing 28), and CSDA (cold shock domain protein A). Co-immunofluorescence and co-immunoprecipitation confirmed these interactions and also interactions with heterochromatin protein-1alpha and acetyl-histone H4 lysine 5. Finally, we investigated the effects of ectopic expression of an RRP1B allelic variant previously associated with improved survival in breast cancer. Gene expression analyses demonstrate that, compared with ectopic expression of wild type RRP1B in HeLa cells, the variant RRP1B differentially modulates various transcription factors controlled by TRIM28 and CSDA. These data suggest that RRP1B, a tumor progression and metastasis susceptibility candidate gene, is potentially a dynamic modulator of transcription and chromatin structure.

B30.2/SPRY domain in tripartite motif-containing 22 is essential for the formation of distinct nuclear bodies

Tripartite motif-containing 22 (TRIM22) is an important antiviral protein that forms distinct nuclear bodies (NB) in many cell types. This study aims to identify functional domains/residues for TRIM22’s nuclear localization and NB formation. Deletion of the really-interesting-new-gene (RING) domain, which is essential for its antiviral property, abolished TRIM22 NB formation. However, mutation of two critical residues Cys15 and Cys18 to alanine in the RING domain, did not affect NB formation notably. Although the deletion of the putative bipartite nuclear localization signal (NLS) abolished TRIM22 localization and NB formation, the B30.2/SplA and ryanodine receptor (SPRY) domain, and residues 491–494 specifically are also essential for nuclear localization and NB formation.

Dynamic localization of tripartite motif-containing 22 in nuclear and nucleolar bodies

Tripartite motif-containing 22 (TRIM22) exhibits antiviral and growth inhibitory properties, but there has been no study on the localization and dynamics of the endogenous TRIM22 protein. We report here that TRIM22 is dramatically induced by progesterone in MDA-MB-231-derived ABC28 cells and T47D cells. This induction was associated with an increase in TRIM22 nuclear bodies (NB), and an even more prominent increase in nucleolar TRIM22 bodies. Distinct endogenous TRIM22 NB were also demonstrated in several other cell lines including MCF7 and HeLa cells. These TRIM22 NB resemble Cajal bodies, co-localized with these structures and co-immunoprecipitated with p80-coilin. However, IFNγ-induced TRIM22 in HeLa and MCF7 cells did not form NB, implying the forms and distribution of TRIM22 are regulated by specific cellular signals. This notion is also supported by the observation that TRIM22 NB undergoes dynamic cell-cycle dependent changes in distribution such that TRIM22 NB started to form in early G0/G1 but became dispersed in the S-phase. In light of its potential antiviral and antitumor properties, the findings here provide an interesting gateway to study the relationship between the different forms and functions of TRIM22.

Microarray analyses of differentially expressed human genes and biological processes in ECV304 cells infected with rubella virus

Changes in mRNA expression levels of ECV304 cells infected with the wild-type rubella strain were analyzed using a microarray system representing 18,716 human genes. Four hundred eighty-seven genes exhibited differential expression levels; 456 of these genes were up-regulated while 31 genes were down-regulated. We identified 53 biological processes that were significantly relevant to the RV-infection. Among these biological processes, 52 were one-gene processes and one was a process involving five genes: IFNA21 (interferon, alpha 21), interferon stimulated exonuclease gene 20 kDa (ISG20), zinc finger protein 175 (ZNF175), tripartite motif-containing 22 (TRIM22), and MX2 [myxovirus (influenza virus) resistance 2 (mouse)]. Except for ZNF175, gene annotation indicated four of these genes encoded interferon or interferon-induced genes. These results suggest that genes relevant to interferon-regulated pathways may be involved in the pathogenesis of rubella.

Tripartite Motif-Containing 29 (TRIM29) Is a Novel Marker for Lymph Node Metastasis in Gastric Cancer

Tripartite motif-containing 29 (TRIM29) belongs to the TRIM protein family, which has unique structural characteristics, including multiple zinc finger motifs and a leucine zipper motif. TRIM29, also known as ataxia telangiectasia group D complementing gene, possesses radiosensitivity suppressor functions. Although TRIM29 has been reported to be underexpressed in prostate and breast cancer, its expression in gastrointestinal cancer has not been studied. By use of real-time reverse transcriptase-polymerase chain reaction, we analyzed TRIM29 mRNA expression status with respect to various clinicopathological parameters in 124 patients with gastric cancer. An immunohistochemical study was also conducted. The expression of TRIM29 was far higher in gastric cancer tumor tissue. Increased TRIM29 mRNA expression was markedly associated with such parameters as histological grade, large tumor size, extent of tumor invasion, and lymph node metastasis. In the TRIM29 high-expression group, it was an independent predictor for lymph node metastasis. Furthermore, patients with high TRIM29 mRNA expression showed a far poorer survival rate than those with low TRIM29 mRNA expression. TRIM29 expression may serve as a good marker of lymph node metastasis in gastric cancer.

In vitro production (IVP) of domestic cat embryos using a modified IVP bovine system

Lipopolysaccharide (LPS) derived from gram negative bacteria cell wall is known to cause ruminal acidosis and/or infectious diseases such as metritis and mastitis which has a significant negative impact on the reproductive performance. This study aimed to investigate the effect of LPS on oocyte maturation and subsequent development in vitro. Ovine cumulus oocyte complexes (COCs) were matured in a medium supplemented with 0 (control), 0.01, 0.1, 1 and 10 μg/mL LPS. Nuclear maturation, cleavage and blastocyst rate, mitochondrial membrane potential (ΔΨm), intracellular reactive oxygen species (ROS) content and changes to the transcript abundance were evaluated. In case of the maturation rate, the percentage of oocytes reaching the MII stage was lower following exposure to 10 μg/mL LPS in comparison to the control group (P < 0.05). Moreover, the blastocyst rate decreased in case of 1 and 10 μg/mL LPS when compared to the control group (P < 0.05). ROS overproduction accompanied by a decreased ΔΨm were recorded in LPS treated oocytes in comparison to the control group (P < 0.05). The 3′ tag digital gene expression profiling method revealed that 7887 genes were expressed while only seven genes exhibited changes in the transcript abundance following exposure to LPS. Tripartite motif containing 25 (TRIM25), Tripartite motif containing 26 (TRIM26), Zona Pellucida glycoprotein 3 (ZP3), Family with sequence similarity 50-member A (FAM50A), Glyoxalate and hydroxy pyruvate reductase (GRHPR), NADH ubiquinase oxireductase subunit A8 (NDUFA8) were down-regulated (P < 0.05), while only Centrin 3 (CETN3) was up-regulated (P < 0.05). Our findings show that LPS has undesirable effects on the maturation competence of ovine oocytes and subsequent embryo development. In addition, the transcriptomic profiling results may shed more light on the molecular mechanisms of LPS-induced infertility in ruminants.