The prolyl 3-hydroxylases P3H2 and P3H3 are novel targets for epigenetic silencing in breast cancer

Shah, Reshma; Smith, Paul; Purdie, Colin A.; Quinlan, P; Baker, Laura A.; Åman, Pierre; Thompson, Alastair; Crook, Tim

doi:10.1038/sj.bjc.6605042

Cited by 54 publications

(55 citation statements)

References 43 publications

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“…5A). The P3H2 gene is reported to be epigenetically silenced in these cells (29). RT-PCR confirmed the absence of P3H2 message in SAOS-2 cells, whereas P3H1, PPIB, CRTAP and P3H3 were robustly expressed (Fig.…”

Section: P3h2 Is Responsible For 3-hydroxylation Of Specific Proline mentioning

confidence: 54%

“…Fig. 5A shows P3H2 protein detected in a lysate of the MB 231 breast cancer cells as expected as these cells are reported to express this gene (29). P3H2 was also detected in CH1.2 cells, a human chondrosarcoma cell line.…”

Section: P3h2 Is Responsible For 3-hydroxylation Of Specific Proline mentioning

confidence: 75%

“…Because RNAi gene silencing was unable to completely knock-out P3H2 activity in RCS-LTC cells, a different approach was pursued. Based on a recent report that P3H2 is epigenetically silenced in some cancer cells (29), cell lysates of cultured human breast cancer, chondrosarcoma, and osteosarcoma cells were screened for P3H2 protein on Western blots. Fig.…”

Section: P3h2 Is Responsible For 3-hydroxylation Of Specific Proline mentioning

confidence: 99%

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A Role for Prolyl 3-Hydroxylase 2 in Post-translational Modification of Fibril-forming Collagens

Fernandes

Farnand

Traeger

et al. 2011

Journal of Biological Chemistry

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The fibrillar collagen types I, II, and V/XI have recently been shown to have partially 3-hydroxylated proline (3Hyp) residues at sites other than the established primary Pro-986 site in the collagen triple helical domain. These sites showed tissue specificity in degree of hydroxylation and a pattern of D-periodic spacing. This suggested a contributory role in fibril supramolecular assembly. The sites in clade A fibrillar ␣1(II), ␣2(V), and ␣1(I) collagen chains share common features with known prolyl 3-hydroxylase 2 (P3H2) substrate sites in ␣1(IV) chains implying a role for this enzyme. We pursued this possibility using the Swarm rat chondrosarcoma cell line (RCS-LTC) found to express high levels of P3H2 mRNA. Mass spectrometry determined that all the additional candidate 3Hyp substrate sites in the pN type II collagen made by these cells were highly hydroxylated. In RNA interference experiments, P3H2 protein synthesis was suppressed coordinately with prolyl 3-hydroxylation at Pro-944, Pro-707, and the C-terminal GPP repeat of the pN␣1(II) chain, but Pro-986 remained fully hydroxylated. Furthermore, when P3H2 expression was turned off, as seen naturally in cultured SAOS-2 osteosarcoma cells, full 3Hyp occupancy at Pro-986 in ␣1(I) chains was unaffected, whereas 3-hydroxylation of residue Pro-944 in the ␣2(V) chain was largely lost, and 3-hydroxylation of Pro-707 in ␣2(V) and ␣2(I) chains were sharply reduced. The results imply that P3H2 has preferred substrate sequences among the classes of 3Hyp sites in clade A collagen chains.Collagen, the major structural protein of vertebrates has evolved a range of post-translational modifications that are essential for triple helix assembly and stability, intermolecular cross-linking, and strength of fibrils and tissue function. Enzymes from the family of 2-oxo-glutarate-dependent dioxygenases are responsible for most of these modifications. For example, prolyl 4-hydroxylase catalyzes proline 4-hydroxylation, a modification necessary for the secondary and tertiary structure of collagen (1, 2), and the lysyl hydroxylase isoenzymes catalyze lysine hydroxylation, a modification essential for the formation of intermolecular cross-links in collagen (2-4).Prolyl 3-hydroxlase 1 (P3H1), 2 another member of the 2-oxo-glutarate-dependent dioxygenase family, catalyzes the post-translational 3-hydroxylation of certain proline residues in fibril-forming collagens. This enzyme was only recently cloned from chicken tissues (5) after being partially purified Ͼ25 years ago (6). P3H1 was identified as the chick homologue of rat leprecan, a glycoprotein, which was first isolated from a rat parietal yolk sac tumor cell line (7). Within the past few years, novel mutations in the human P3H1 gene and genes encoding P3H1-associated proteins were shown to cause recessive forms of osteogenesis imperfecta (8 -11). By mass spectrometry, we and other investigators have shown that Pro-986 in the ␣1(I) collagen chain of bone and skin from such patients can be severely under-3-hydroxyated. The P3H1 enz...

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Section: P3h2 Is Responsible For 3-hydroxylation Of Specific Proline mentioning

confidence: 54%

Section: P3h2 Is Responsible For 3-hydroxylation Of Specific Proline mentioning

confidence: 75%

Section: P3h2 Is Responsible For 3-hydroxylation Of Specific Proline mentioning

confidence: 99%

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A Role for Prolyl 3-Hydroxylase 2 in Post-translational Modification of Fibril-forming Collagens

Fernandes

Farnand

Traeger

et al. 2011

Journal of Biological Chemistry

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“…P3Hs are clearly expressed in 293-HEK cells at levels similar to those of a cell line previously shown positive for expression of all three enzymes (43). CRTAP and peptidyl prolyl cis-trans isomerase B expression levels were readily detectable as well.…”

Section: Hydroxylated Residues and Glycosylated Hyl Residues Of Humanmentioning

confidence: 67%

“…cDNA was synthesized from 1 g of RNA using the SuperScript II Reverse Transcriptase kit (Invitrogen) with random hexamers. Subsequently, PCR was performed for 35 cycles with primer sets for human prolyl 3-hydroxylase 1 (P3H1), P3H2, P3H3, cartilage-associated protein (CRTAP), and PPIB previously described by Fernandes et al (31) and for GAPDH previously described by Shah et al (43).…”

Section: ␣1(v)mentioning

confidence: 99%

Comprehensive Mass Spectrometric Mapping of the Hydroxylated Amino Acid residues of the α1(V) Collagen Chain

Yang

Park

Davis

et al. 2012

Journal of Biological Chemistry

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Background: ␣1(V) is an extensively modified collagen chain important in disease. Results: Comprehensive mapping of ␣1(V) post-translational modifications reveals unexpectedly large numbers of X-position hydroxyprolines in Gly-X-Y amino acid triplets. Conclusion:The unexpected abundance of X-position hydroxyprolines suggests a mechanism for differential modification of collagen properties. Significance: Positions, numbers, and occupancy of modified sites can provide insights into ␣1(V) biological properties.

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The role of network‐forming collagens in cancer progression

Zhang

Liu

Zhang

et al. 2022

Intl Journal of Cancer

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Collagens are the main components of extracellular matrix in the tumor microenvironment. Both fibrillar and nonfibrillar collagens are involved in tumor progression. The nonfibrillar network‐forming collagens such as type IV and type VIII collagens are frequently overexpressed in various types of human cancers, which promotes tumor cell proliferation, adhesion, invasion, metastasis and angiogenesis. Studies on the roles of these collagens have shed light on the mechanisms underpinning the effects of this protein family. Future research has to explicit the role of network‐forming collagens with respect to cancer progression and treatment. Herein, we review the regulation of network‐forming collagens expression in cancer; the roles of network‐forming collagens in tumor invasion, metastasis and angiogenesis; and the clinical significance of network‐forming collagens expression in cancer patients.

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The prolyl 3-hydroxylases P3H2 and P3H3 are novel targets for epigenetic silencing in breast cancer

Cited by 54 publications

References 43 publications

A Role for Prolyl 3-Hydroxylase 2 in Post-translational Modification of Fibril-forming Collagens

A Role for Prolyl 3-Hydroxylase 2 in Post-translational Modification of Fibril-forming Collagens

Comprehensive Mass Spectrometric Mapping of the Hydroxylated Amino Acid residues of the α1(V) Collagen Chain

The role of network‐forming collagens in cancer progression

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