The mitotically phosphorylated form of the transcription termination factor TTF-1 is associated with the repressed rDNA transcription machinery

1999 ◽  
Vol 112 (19) ◽  
pp. 3259-3268 ◽  
Author(s):  
V. Sirri ◽  
P. Roussel ◽  
D. Hernandez-Verdun
Keyword(s):  
Transcription Termination ◽  
Nucleolar Organizer ◽  
Ribosomal Gene ◽  
Nucleolar Organizer Regions ◽  
Rdna Transcription ◽  
Mitotic Chromosomes ◽  
Termination Factor ◽  
Transcription Machinery ◽  
Phosphorylated Form ◽  
Transcription Termination Factor

The transcription termination factor TTF-1 exerts two functions in ribosomal gene (rDNA) transcription: facilitating initiation and mediating termination of transcription. Using HeLa cells, we show that TTF-1 protein is colocalized with the active transcription machinery in the nucleolus and also with the inactive machinery present in certain mitotic nucleolar organizer regions (NORs) when rDNA transcription is repressed. We also show that TTF-1 is specifically phosphorylated during mitosis in a manner dependent on the cdc2-cyclin B kinase pathway and on an okadaic acid-sensitive phosphatase. Interestingly, the mitotically phosphorylated form of TTF-1 appearing at the G(2)/M transition phase was more easily solubilized than was the interphase form. This indicates that the chromatin-binding affinity of TTF-1 appears to be different in mitotic chromosomes compared to the interphase nucleolus. Correlated with this, the other DNA-binding factor, UBF, which interferes with chromatin conformation in the rDNA promoter, was more strongly bound to rDNA during mitosis than at interphase. The reorganization of the mitotic rDNA promoter might be induced by phosphorylation of certain components of the rDNA transcription machinery and participate in silencing of rDNA during mitosis.

A role for upstream binding factor in organizing ribosomal gene chromatin

2006 ◽  
Vol 73 ◽  
pp. 77-84 ◽  
Author(s):  
Jane E. Wright ◽  
Christine Mais ◽  
José-Luis Prieto ◽  
Brian McStay
Keyword(s):  
Nucleolar Organizer ◽  
Ribosomal Gene ◽  
Rna Polymerase I ◽  
Nucleolar Organizer Regions ◽  
Binding Factor ◽  
Upstream Binding Factor ◽  
Acrocentric Chromosomes ◽  
Polymerase I ◽  
Secondary Constrictions ◽  
Active Nors

Human ribosomal genes are located in NORs (nucleolar organizer regions) on the short arms of acrocentric chromosomes. During metaphase, previously active NORs appear as prominent chromosomal features termed secondary constrictions, which are achromatic in chromosome banding and positive in silver staining. The architectural RNA polymerase I transcription factor UBF (upstream binding factor) binds extensively across the ribosomal gene repeat throughout the cell cycle. Evidence that UBF underpins NOR structure is provided by an examination of cell lines in which large arrays of a heterologous UBF binding sequences are integrated at ectopic sites on human chromosomes. These arrays efficiently recruit UBF even to sites outside the nucleolus, and during metaphase form novel silver-stainable secondary constrictions, termed pseudo-NORs, that are morphologically similar to NORs.

Structure and variability of nucleolar organizer regions in Parodontidae fish

1984 ◽  
Vol 26 (5) ◽  
pp. 564-568 ◽  
Author(s):  
Orlando Moreira-Filho ◽  
Luiz Antonio Carlos Bertollo ◽  
Pedro Manoel Galetti Jr.
Keyword(s):  
Chromosome Pair ◽  
Constitutive Heterochromatin ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions ◽  
Mitotic Chromosomes ◽  
Homozygous Condition ◽  
Nucleolar Organizing Regions ◽  
Nucleolar Chromosome

Nucleolar organizer regions (NORs) were studied in mitotic chromosomes of four species of fish of family Parodontidae: Parodon tortuosus, Apareiodon affinis, Apareiodon ibitiensis, and Apareiodon piracicabae. All four species exhibited only a single nucleolar chromosome pair in their karyotypes. Intraspecific differences were observed in the size of these chromosomes; however, these were not very clear for A. affinis and A. piracicabae, Apareiodon piracicabae exhibited two clearly visible NORs in each of the nucleolar chromosomes, which was the only configuration practically found in this species. This trait therefore predominates in a homozygous condition in the population investigated. Regions of constitutive heterochromatin adjacent to the two NORs were detected. Possible mechanisms that may have originated the two NORs are discussed.Key words: nucleolar organizing regions, fish.

Bacteriophage T4 Alc protein: A transcription termination factor sensing local modification of DNA

Cell ◽  
1993 ◽  
Vol 75 (1) ◽  
pp. 147-154 ◽  
Author(s):  
Mikhail Kashlev ◽  
Evgeny Nudler ◽  
Alex Goldfarb ◽  
Terry White ◽  
Elizabeth Kutter
Keyword(s):  
Transcription Termination ◽  
Bacteriophage T4 ◽  
Protein A ◽  
Termination Factor ◽  
Transcription Termination Factor ◽  
Local Modification

Mitosis-specific phosphorylation of nucleolin by p34cdc2 protein kinase

1990 ◽  
Vol 10 (7) ◽  
pp. 3607-3618
Author(s):  
P Belenguer ◽  
M Caizergues-Ferrer ◽  
J C Labbé ◽  
M Dorée ◽  
F Amalric
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Control ◽  
Nucleolar Organizer ◽  
Serine Phosphorylation ◽  
Nucleolar Organizer Regions ◽  
Rdna Transcription ◽  
Amino Terminal ◽  
Nucleolar Chromatin

Nucleolin is a ubiquitous multifunctional protein involved in preribosome assembly and associated with both nucleolar chromatin in interphase and nucleolar organizer regions on metaphasic chromosomes in mitosis. Extensive nucleolin phosphorylation by a casein kinase (CKII) occurs on serine in growing cells. Here we report that while CKII phosphorylation is achieved in interphase, threonine phosphorylation occurs during mitosis. We provide evidence that this type of in vivo phosphorylation involves a mammalian homolog of the cell cycle control Cdc2 kinase. In vitro M-phase H1 kinase from starfish oocytes phosphorylated threonines in a TPXK motif present nine times in the amino-terminal part of the protein. The same sites which matched the p34cdc2 consensus phosphorylation sequence were used in vivo during mitosis. We propose that successive Cdc2 and CKII phosphorylation could modulate nucleolin function in controlling cell cycle-dependent nucleolar function and organization. Our results, along with previous studies, suggest that while serine phosphorylation is related to nucleolin function in the control of rDNA transcription, threonine phosphorylation is linked to mitotic reorganization of nucleolar chromatin.

When rDNA transcription is arrested during mitosis, UBF is still associated with non-condensed rDNA

1997 ◽  
Vol 110 (19) ◽  
pp. 2429-2440 ◽  
Author(s):  
J. Gebrane-Younes ◽  
N. Fomproix ◽  
D. Hernandez-Verdun
Keyword(s):  
Secondary Constriction ◽  
Three Dimensional ◽  
Ribosomal Gene ◽  
Rdna Transcription ◽  
Transcription Machinery ◽  
Late Anaphase ◽  
Early Anaphase ◽  
Dimensional Distribution ◽  
Open Question ◽  
Kinetics Of

The mechanisms that control inactivation of ribosomal gene (rDNA) transcription during mitosis is still an open question. To investigate this fundamental question, the precise timing of mitotic arrest was established. In PtK1 cells, rDNA transcription was still active in prophase, stopped in prometaphase until early anaphase, and activated in late anaphase. Because rDNA transcription can still occur in prophase and late anaphase chromosomes, the kinetics of rDNA condensation during mitosis was questioned. The conformation of the rDNA was analyzed by electron microscopy from the G2/M transition to late anaphase in the secondary constriction, the chromosome regions where the rDNAs are clustered. Whether at transcribing or non-transcribing stages, non-condensed rDNA was observed in addition to axial condensed rDNA. Thus, the persistence of this non-condensed rDNA during inactive transcription argues in favor of the fact that mitotic inactivation is not the consequence of rDNA condensation. Analysis of the three-dimensional distribution of the rDNA transcription factor, UBF, revealed that it was similar at each stage of mitosis in the secondary constriction. In addition, the colocalization of UBF with non-condensed rDNA was demonstrated. This is the first visual evidence of the association of UBF with non-condensed rDNA. As we previously reported that the rDNA transcription machinery remained assembled during mitosis, the colocalization of rDNA fibers with UBF argues in favor of the association of the transcription machinery with certain rDNA copies even in the absence of transcription. If this hypothesis is correct, it can be assumed that condensation of rDNA as well as dissociation of the transcription machinery from rDNA cannot explain the arrest of rDNA transcription during mitosis. It is proposed that modifications of the transcription machinery occurring in prometaphase could explain the arrest of transcription, while reverse modifications in late anaphase could explain activation.

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