The nanovirus-encoded Clink protein affects plant cell cycle regulation through interaction with the retinoblastoma-related protein.
Nanoviruses, multicomponent single-stranded DNA plant viruses, encode a unique cell cycle link protein, Clink, that interacts with retinoblastoma-related proteins (RBR). We have established transgenic Arabidopsis thaliana lines that conditionally express Clink or a Clink variant deficient in RBR binding. By controlled induction of Clink expression, we demonstrated the capacity of the Clink protein to alter RBR function in vivo. We showed that transcription of both S-phase-specific and G2/M-phase-specific genes was up-regulated depending on the RBR-binding proficiency of Clink. Concomitantly, ploidy levels increased in a substantial fraction of leaf cell nuclei. Also, leaf epidermis cells of transgenic plants producing Clink were smaller and more numerous, indicating additional cell divisions in this tissue. Furthermore, cytogenetic analyses following induction of Clink expression in mature leaves revealed the presence of metaphasic and anaphasic nuclei, clear evidence that Clink-mediated RBR inactivation is sufficient to induce quiescent cells to reenter cell cycle progression and, for at least a fraction of them, to pass through mitosis. Expression of Clink had no effect on genes transcribed by RNA polymerases I and III, suggesting that, in contrast to its mammalian homologue, A. thaliana RBR is not involved in the repression of polymerase I and polymerase III transcription. The results of these in vivo analyses firmly establish Clink as a member of the diverse class of multifunctional cell cycle modulator proteins encoded by small DNA viruses.
Lageix
Sébastien
S
CNRS UMR 6547 BIOMOVE, Université Blaise Pascal, 24 Avenue des Landais, 63177 Aubière Cedex, France.
Catrice
Olivier
O
Deragon
Jean-Marc
JM
Gronenborn
Bruno
B
Pélissier
Thierry
T
Ramírez
Bertha Cecilia
BC
D017360:Arabidopsis / Q000821:virology*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology*; D002455:Cell Division; D002467:Cell Nucleus / Q000235:genetics; D016195:G2 Phase; D015870:Gene Expression; D005786:Gene Expression Regulation; D025682:Nanovirus / Q000502:physiology*; D030821:Plants, Genetically Modified; D011123:Polyploidy; D016196:S Phase; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D029681:Arabidopsis Proteins; C487568:RBR1 protein, Arabidopsis; D014764:Viral Proteins