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The concept of the CCN protein family revisited: a centralized coordination network

Journal of Cell Communication and Signaling volume 12, pages 3–12 (2018)Cite this article

Abstract

The wide array of biological properties attributed to the CCN family of proteins (Perbal in Lancet 363(9402):62–64, 2004) led me to reconsider the possible relationship and roles that these proteins may play as a team, instead of acting on their own as individual regulators in various signaling pathways. The dynamic model which I present in this review stems from the contribution of the biological properties that we established for CCN3, one of the three founding members of the CCN family, which was identified by our group as the first CCN protein showing growth inhibitory properties (1992), expressed mainly in quiescent cells (1996), and showing anti-tumor activities in several cellular models both ex vivo and in vivo. At the present time CCN3 is the only member of the family that has been reported to negatively act on the progression of the cell cycle. The unique dual localisation of CCN3 in the nucleus and outside cells, either at the membrane or in the extracellular matrix, that I first established in 1999, and that now appears to be shared by several other CCN proteins, is a unique essential feature which can no longer be ignored. Based on the structural and functional properties of CCN3, shared by most of the CCN family members, I propose an « all in one » concept in which CCN proteins are team members with specific functions that are aimed at the same goal. This model accounts both for the functional specificity of the various CCN proteins, their sequential and opposite or complementary effects in various biological context, and for the biological consequences of their physical interaction and biological cross-regulation.

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Notes

  1. Several groups, including mine, have in the past used recombinant proteins and individual domains produced in bacterial or baculoviral expression systems which proved to show biological activities exhibited by the native proteins (for CCN3 see Perbal et al. 1999 and Lazar et al. 2007). Unfortunately these proteins do not show the whole panel of post transcriptional modifications that have been suspected for a long time to be essential for native eucaryotic proteins to interact with their physiological partners.

  2. On many occasions, since the early 1990s, efforts to attract colleagues specialized in physicochemical protein analysis to work on CCN proteins, proved unsuccessful and the feeling developed that proteins showing such a complex array of biological properties were not good candidates for this type of studies.

  3. The authors quote CCN3 was amplified from cDNA (a kind gift from Dr. Paul Kemp). Hopefully this clone was not the one used in previous studies from Kemp’s group (Ellis et al., 2000) who claimed that their « data indicate that « NOV is not an effective inhibitor of proliferation, at least in vitro ». In our hands the human and chicken recombinant CCN3 proteins that we purified, and that were also used by others, showed both anti-proliferative and anti-tumor activities.

  4. Perbal B, Lau L, Lyons K, Kubota S, Yeger H, Fisher G.(2016) Report on the 8th international workshop on the CCN family of genes - Nice November 3–8, 2015. J Cell Commun Signal. 10:77–86.

  5. « CCN proteins might act as multipotent matchmakers, allowing the scaffolding of a complex network of regulatory proteins responsible for the regulation of several fundamental functions during normal life, development, and probably death. The identification of other proteins potentially interacting with NOV has established that each of the four constitutive domains of the native protein is required alone or in combination with others to promote the interactions » (Perbal 2001)

  6. I wish to take this opportunity to point out an erroneous affirmation that was published regarding the biological properties of CCN3 and that conveyed wrong concepts. Although our results had clearly established that CCN3 inhibited cell growth on plastic, Ellis et al. (2000) quoted nine years later in « the data suggest that it [CCN3] is an inhibitor of cell proliferation, first demonstrated for chick embryo fibroblasts (CEFs) grown in soft agar » This affirmation is erroneous: i) CEF do not grow in agar unless they are transformed, and ii) our experimental conditions used primary CEF infected by retroviral expression vectors expressing CCN3 either in the sense or in the antisense orientations. When cells were plated, only those which do not expressed CCN3 could grow. Those which expressed CCN3 did not grow. The statement of Ellis et al. would imply that CCN3 avoids CEF-infected cells cloning without anchorage.

  7. « The discovery of nov had three immediate implications: (1) The structural relation that had been reported between cyr61 and ctgf was extended to another gene, thereby leading to the concept of a new family of genes (a family is usually defined as a group of at least three subjects sharing common characteristics). (2) nov provided the first example of aberrant expression of a CCN gene being associated with tumour development, therefore indicating that the biological properties of this new class of growth regulators extended beyond the normal response to growth stimulation. (3) The growth inhibitory effect of nov on chicken embryonic fibroblasts (CEFs) also provided the first evidence that the CCN family of genes contains negative regulators and suggested that nov might counterbalance the stimulatory effects of the immediate early CCN proteins. » (Perbal 2001)

  8. « The rCop-1 gene was found to be expressed only when primary REFs and MEFs began to age or became senescent during passage in culture » The authors also proposed « Since the rCOP-1 protein lacks the C-terminal module of the canonical CCN family members, one can expect it to act as a « dominant negative » effector in the homo and heterotropic multimerization of CCN proteins, either with other members of the CCN family or with proteins which do not belong to the CCN family but physically interact with CCN family members » (Zhang et al. 1998 )

  9. In their manuscript, the authors claim that « rCOP-1 represents a new class of CCN proteins that have functions opposing those of previously indentified members »…. The authors did not seem to fully appreciate that the anti proliferative activity of NOV was sufficient enough to distinguish it from other CCN proteins (see Perbal 2013)

  10. Surprisingly, 11 years after we had experimentally demonstrated that CCN3 was an antiproliferative factor in several different cell types, the authors of this manuscript falsely reported that « Although it is hypothesized that CCN3 may be a negative regulator of proliferation, there is no direct evidence to date supporting this function. » Furthermore, they also claimed that « recombinant CCN3 has been unable thus far to inhibit proliferation of cells in culture, including VSMCs » and added that their observations suggested « the intriguing possibility that CCN5 may represent a naturally occurring antagonist to the 4-domain members of the CCN family »!

  11. « A clue to understanding the discrepancies observed for the expression of the CCN genes in cancer cells might come from the multimodular structure of the CCN proteins and their potential ability to interact sequentially or simultaneously with different partners, the bioavailability of which might be a key element in the manifestation of the positive or negative effects attributed to the CCN proteins »…    « CCN proteins might act as multipotent matchmakers, allowing the scaffolding of a complex network of regulatory proteins responsible for the regulation of several fundamental functions during normal life, development, and probably death. The identification of other proteins potentially interacting with NOV has established that each of the four constitutive domains of the native protein is required alone or in combination with others to promote the interactions. ». B. Perbal (2001)

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Acknowledgements

I am deeply grateful to Dr. Herman Yeger for his critical review of the manuscript and to Annick for her help and support.

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Affiliations

  1. Université Côte d’Azur, CNRS, GREDEG, International CCN Society, Nice, France

    Bernard Perbal

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  1. Bernard Perbal
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Correspondence to Bernard Perbal.

Additional information

This review is based on the presentation that I delivered as an introduction to the 9th International Workshop on the CCN family of genes held in Saint-Malo France (Nov 2-7th, 2017)

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Perbal, B. The concept of the CCN protein family revisited: a centralized coordination network. J. Cell Commun. Signal. 12, 3–12 (2018). https://doi.org/10.1007/s12079-018-0455-5

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Keywords

  • Signaling coordination
  • Development
  • Angiogenesis
  • Differentiation
  • Inflammation
  • Growth control
  • Nuclear proteins
  • Matricellular proteins
  • Extracellular matrix (ECM)
  • Tumorigenic potential
  • Regulation of cell cycle
  • Thrombospondin
  • Von Willebrand factor
  • Growth factors
  • IG Fbinding proteins (IGFBPs)