Tyrosine phosphorylation controls PCNA function through protein stability

Wang, Shao‐Chun; Nakajima, Yusuke; Yu, Yung Luen; Xia, Weiya; Chen, Chun‐Te; Yang, Cheng Chieh; McIntush, Eric W.; Li, Long Yuan; Hawke, David H.; Kobayashi, Ryûji; Hung, Mien‐Chie

doi:10.1038/ncb1501

Cited by 277 publications

(282 citation statements)

References 45 publications

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“…S2A). To confirm further the nuclear location of EGFR/RHA complexes, sequential photosections of a nucleus were examined, and EGFR/RHA complexes were clearly detected in middle sections (i.e., planes [11][12][13][14] in both MDA-MB-468 and HeLa cells (Fig. S2 B and C).…”

Section: Resultsmentioning

confidence: 99%

“…Nuclear EGFR has been demonstrated to contribute to cancer cell resistance to cetuximab and radiation treatment (5,6) and to be negatively correlated with overall survival of patients with multiple cancer types (7)(8)(9)(10)(11). Moreover, nuclear EGFR interacts with signal transducer and activator of transcription 3 (STAT3), signal transducer and activator of transcription 5A (STAT5A), E2F1, DNA-dependent protein kinase (DNA-PK), and proliferating cell nuclear antigen (PCNA) and plays important roles in cell transformation, proliferation, and DNA repair and replication (12)(13)(14)(15)(16). Nuclear EGFR regulates gene expression by binding to an AT-rich sequence (ATRS) of the gene's promoter (13,16,17).…”

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confidence: 99%

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RNA helicase A is a DNA-binding partner for EGFR-mediated transcriptional activation in the nucleus

Huo

Wang

Xia

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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EGF induces the translocation of EGF receptor (EGFR) from the cell surface to the nucleus where EGFR activates gene transcription through its binding to an AT-rich sequence (ATRS) of the target gene promoter. However, how EGFR, without a DNA-binding domain, can bind to the gene promoter is unclear. In the present study, we show that RNA helicase A (RHA) is an important mediator for EGFRinduced gene transactivation. EGF stimulates the interaction of EGFR with RHA in the nucleus of cancer cells. The EGFR/RHA complex then associates with the target gene promoter through binding of RHA to the ATRS of the target gene promoter to activate its transcription. Knockdown of RHA expression in cancer cells abrogates the binding of EGFR to the target gene promoter, thereby reducing EGF/EGFR-induced gene expression. In addition, interruption of EGFR-RHA interaction decreases the EGFR-induced promoter activity. Consistently, we observed a positive correlation of the nuclear expression of EGFR, RHA, and cyclin D1 in human breast cancer samples. These results indicate that RHA is a DNA-binding partner for EGFR-mediated transcriptional activation in the nucleus.cyclin D1 | nuclear translocation | inducible nitric oxide synthase | transcription C ell surface EGF receptor (EGFR) has been shown to be localized in the nucleus (1-4). Nuclear EGFR has been demonstrated to contribute to cancer cell resistance to cetuximab and radiation treatment (5, 6) and to be negatively correlated with overall survival of patients with multiple cancer types (7-11). Moreover, nuclear EGFR interacts with signal transducer and activator of transcription 3 (STAT3), signal transducer and activator of transcription 5A (STAT5A), E2F1, DNA-dependent protein kinase (DNA-PK), and proliferating cell nuclear antigen (PCNA) and plays important roles in cell transformation, proliferation, and DNA repair and replication (12-16). Nuclear EGFR regulates gene expression by binding to an AT-rich sequence (ATRS) of the gene's promoter (13,16,17). Additionally, a recent unbiased protein-DNA interactome study indicates that EGFR is a DNAbinding protein (18). However, EGFR does not contain a DNAbinding domain, and evidence supporting direct binding of EGFR to the specific DNA sequence is lacking. Thus, identifying the DNA-binding partner for EGFR is crucial for understanding how EGFR regulates gene transcription in the nucleus.RNA helicase A (RHA), the human homolog of Drosophila maleless (MLE) that increases the transcription of male X-linked genes (19), is a multifunctional protein and is conserved in Drosophila and mammals (20)(21)(22). RHA belongs to the aspartateglutamate-alanine-aspartate (DEAD) box family of proteins and has the ability to bind to RNA and DNA (23,24). RHA regulates gene transcription by interacting with transcription factors (22) or by binding directly to the target gene promoter (25). Moreover, Drosophila MLE activates rox2 transcription by binding to an ATrich region of the gene promoter (26). Interestingly, this AT-rich region contains the previo...

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

RNA helicase A is a DNA-binding partner for EGFR-mediated transcriptional activation in the nucleus

Huo

Wang

Xia

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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“…Phospho-PCNA is correlated with nuclear EGFR in human breast cancer tumor samples and has a strong prognostic value for breast cancer patients survival. Nuclear EGFR is also positively correlated with the expression of PCNA in oral squamous cancer [28].…”

Section: Egfr and The Hr Mechanismmentioning

confidence: 86%

Role of epidermal growth factor receptor in DNA damage repair

Yang

Tao³

et al. 2011

Chin. Sci. Bull.

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Many human tumor cells are characterized by overexpression or mutation of epidermal growth factor receptor (EGFR). Emerging evidence indicates that EGFR, as well as some of its downstream components, can translocate to the nucleus and play roles in transcriptional regulation, signaling conduction and repair of DNA double strands breaks (DSBs). EGFR in its nuclear manifestation promotes DSB repair by interacting with proteins including DNA-PK, ATM, Rad51 and BRCA1, involved in DSB repair, via the PI3K-Akt and Ras-Raf-MAPK pathways. DNA damage repair in tumor cells is emerging as an attractive target in radiotherapy and chemotherapy. Interruption of EGFR functions, or those of its downstream components, presents a promising strategy for confounding DNA damage repair in tumor cells. The epidermal growth factor receptor (EGFR) belongs to the Type I subfamily of receptor tyrosine kinases (RTKs), which includes four closely related receptors: Her-1/ErbB1, Her-2/Neu/ErbB2, Her-3/ErbB3 and Her-4/ErbB4. EGFR is a 170-kD transmembrane glycoprotein composed of 1186 aa residues. Structurally EGFR has an extracellular ligandbinding domain, a transmembrane domain, and an intracellular tyrosine kinase domain (TKD). The function and downstream signaling of EGFR as a cell surface receptor have been documented by a substantial body of literature. It is becoming abundantly clear, however, that aside from its duties in the cell membrane, EGFR and some of its downstream components translocate to the nucleus where they have various regulatory functions, such as transcriptional regulation and repair processes. Among these, the function of nuclear EGFR as a regulator of double strands breaks (DSBs) is of particular importance [1]. In this article, we review the current knowledge regarding EGFR modulation of DNA damage repair.

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“…Several reports have suggested the existence of PCNA modifications, 9,[16][17][18] ranging from phosphorylation, 10,19,20 not confirmed by others' data, 21,22 to acetylation 22 and, recently, to methyl esterification. 23 Additionally, PCNA mono and poly-ubiquitylation in response to DNA damage and sumoylation in the absence of damage was described.…”

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confidence: 95%

Human hepatocellular carcinoma expresses specific PCNA isoforms: an in vivo and in vitro evaluation

Venturi

Piaz

Giovannini

et al. 2008

Laboratory Investigation

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Proliferating cell nuclear antigen (PCNA) is a 36 kDa protein involved in several cellular mechanisms, including DNA synthesis and repair, cell cycle regulation and apoptosis. An alteration in PCNA structure might contribute to DNA-damage accumulation in cancer cells. This study was aimed to evaluate the PCNA pattern of expression, in terms of aggregation status, isoforms and post-translational modifications, in human hepatocellular carcinoma (HCC) and cirrhosis as well as in HCC cell lines. Twelve HCCs and surrounding cirrhotic tissues were analysed, along with HepG2, Hep3B and SNU-398 cell lines. Normal liver specimens and cirrhosis without HCC were included as controls. Both DNA-bound and DNA-unbound PCNA fractions were analysed, and PCNA pattern of expression was displayed on two-dimensional gel electrophoresis followed by western blot. Results were confirmed by mass spectrometry. To compare HCCs vs surrounding tissues, immunolabelling and immunostaining were performed. In 6 of 12 HCCs and in cell lines, we found three major PCNA acidic forms, corresponding to monomers, probably dimers and trimers, and a basic isoform. In the six remaining HCCs, only a PCNA acidic form associated with multiple basic isoforms was detected. Importantly, the PCNA basic form was not found in cirrhotic tissues. To clarify the nature of the detected PCNA isoforms, ubiquitin-specific immunoblotting as well as phosphatase treatment were employed. A PCNA-ubiquitylated form in cell lines and PCNA-phosphorylated isoforms in 6 of 12 HCCs were detected. Finally, in the DNA-bound fraction we detected only an acidic PCNA monomeric form. We conclude that human hepatocellular carcinoma expresses specific PCNA isoforms compared to those found in cirrhosis, implicating a role for PCNA functional alterations in hepatocarcinogenesis.

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Tyrosine phosphorylation controls PCNA function through protein stability

Cited by 277 publications

References 45 publications

RNA helicase A is a DNA-binding partner for EGFR-mediated transcriptional activation in the nucleus

RNA helicase A is a DNA-binding partner for EGFR-mediated transcriptional activation in the nucleus

Role of epidermal growth factor receptor in DNA damage repair

Human hepatocellular carcinoma expresses specific PCNA isoforms: an in vivo and in vitro evaluation

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