Surface residue mutations of the PML RING finger domain alter the formation of nuclear matrix-associated PML bodies.
The human protein PML, was first identified as part of a fusion protein with retinoic acid receptor alpha as found in the chromosomal translocation which gives rise to acute promyelocytic leukaemia. PML is normally localised to large matrix-associated nuclear domains (known as ND10, Kr bodies, PODS or PML NBs) which comprise several multi-protein complexes. Within the PML protein, there are a number of identified zinc-binding domains, one of which called the RING finger is found in a large family of diverse and unrelated proteins. Here, we report the effect of site-directed mutations within the context of the whole PML protein, of amino acids found on the surface of the PML RING finger domain and PML NB formation in vivo. Mutations of a small region of the RING finger domain surface affect the size and numbers of PML NBs in a mouse fibroblast expression assay, resulting in fewer but larger exogenous PML NBs. Mutations of other surface RING residues, however, do not affect exogenous PML NB formation. Furthermore, all of the PML RING mutants co-localise to both endogenous and exogenous wild-type PML NBs. These data identify a specific region of the PML RING finger domain which is directly involved in correct PML NB formation. They also provide evidence to suggest that the PML RING finger is involved in mediating PML-PML oligomeric interactions, as part of a mechanism leading to the assembly of the PML NB complex.
Boddy
M N
MN
The Scripps Research Institute, Department of Molecular Biology, La Jolla, CA 92037, USA.
Duprez
E
E
Borden
K L
KL
Freemont
P S
PS
D016475:3T3 Cells / Q000502:physiology; D000818:Animals; D018698:Glutamic Acid; D006367:HeLa Cells; D006801:Humans; D008239:Lysine; D051379:Mice; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D009363:Neoplasm Proteins; D015530:Nuclear Matrix / Q000737:chemistry*; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D017434:Protein Structure, Tertiary; D055672:Static Electricity; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics*; D014162:Transfection; D025521:Tumor Suppressor Proteins; D016335:Zinc Fingers / Q000235:genetics*
D009363:Neoplasm Proteins; D009687:Nuclear Proteins; C513293:Pml protein, mouse; D000071156:Promyelocytic Leukemia Protein; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; D018698:Glutamic Acid; D008239:Lysine