The nucleus of axenically grownDictyostelium discoideum: Studies on its division cycle, isolation and conformation

Maclean, Norman; Garside, K.; Bradley, M. C.; Wood, Clive

doi:10.1007/bf01946648

Cited by 10 publications

(5 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The pellets were resuspended in lysis buffer and re-centrifuged, yielding the P3 and P100 (where P100 is the particulate cell fraction equivalent to the pellet formed by centrifugation at 100,000 ϫ g for 1 h) fractions, respectively. Nuclei were prepared using either Nonidet P-40 or digitonin as described (21). The S100 supernatant (800 ml) was pumped onto a 450-ml DEAE-Sepharose Fast Flow column equilibrated in buffer A and eluted with the same buffer (17).…”

Section: Glcnac-transferase Purificationmentioning

confidence: 99%

Identification of a UDP-GlcNAc:Skp1-Hydroxyproline GlcNAc-transferase in the Cytoplasm of Dictyostelium

Teng-umnuay¹,

Wel

West

1999

Journal of Biological Chemistry

View full text Add to dashboard Cite

Skp1 is a cytoplasmic and nuclear protein required for the ubiquitination of cell cycle regulatory proteins and transcriptional factors. In Dictyostelium, Skp1 is modified by a linear pentasaccharide, Gal␣1-6Gal␣1-Fuc␣1-2Gal␤1-3GlcNAc, attached to a hydroxyproline (HyPro) residue at position 143. To study the formation of the GlcNAc-HyPro linkage, an assay was developed for the transfer of Skp1 belongs to the SCF complex that is involved in the ubiquitination of cell cycle and other regulatory proteins including transcriptional factors (1-4). In Dictyostelium discoideum, Skp1 is encoded by two similar genes, fpa1 and fpa2 (5), and is found in both the cytoplasm and the nucleus based on immunofluorescence localization.

show abstract

Section: Glcnac-transferase Purificationmentioning

confidence: 99%

Identification of a UDP-GlcNAc:Skp1-Hydroxyproline GlcNAc-transferase in the Cytoplasm of Dictyostelium

Teng-umnuay¹,

Wel

West

1999

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Crude nuclei were isolated by the method of Maclean et al (1984), with minor modifications. Growing amoebae, and amoebae that had been heat shocked, were washed once in ice-cold 20 mM phosphate buffer, pH 6.4, and then resuspended in ice-cold nuclear buffer (50 mM HEPES, pH 7.5, 100 mM KCl, 5 mM magnesium acetate 10% sucrose, 1 mM phenylmethylsulfonyl fluoride, 0.1% Triton X-100) at 110 8 cells/ml.…”

Section: Methodsmentioning

confidence: 99%

“…Nuclei of AX-2 cells typically contain two to four nucleoli (Maclean et al 1984). Observations in the parental strain, NC-4, suggest that the nucleolus is a single multilobed structure .…”

Section: Hsp32 Is a Nucleolar Proteinmentioning

confidence: 99%

Dictyostelium discoideum Hsp32 is a resident nucleolar heat-shock protein

Moerman

Klein

1998

Chromosoma

View full text Add to dashboard Cite

Hsp32 is a small shock protein in Dictyostelium discoideum that is expressed in growing cells in the absence of heat shock. Here we show that Hsp32 is an Ag-NOR-staining protein capable of binding DNA with high affinity. Hsp32 is also shown to be a resident nucleolar protein both under normal growth conditions and during heat stress. In unstressed cells, Hsp32 localizes to the nucleolar periphery in a pattern reminiscent of the rDNA in this organism. During the first several hours of heat shock, the peripheral localization of Hsp32 is not altered, although rDNA transcription is arrested. Prolonged heat shock causes a condensation of the rDNA. Under these conditions, Hsp32 is no longer predominantly associated with the rDNA, but is instead distributed over the entire nucleolus. Hsp32 therefore retains ist nucleolar localization under prolonged heat shock conditions by associating with nucleolar components other than the rDNA or rRNA.

show abstract

“…Early research revealed that the structural features of this social amoebozoan’s nucleoli differ from the classic nucleolar organization. Rather than localizing within the nucleoplasm, they exist as two to four dense patches that are tightly adhered to the inner nuclear envelope (Figure 1) [7,8,9,10,11]. What’s more, they are neither bipartite nor tripartite.…”

Section: The Dictyostelium Nucleolusmentioning

confidence: 99%

Proteins of the Nucleolus of Dictyostelium discoideum: Nucleolar Compartmentalization, Targeting Sequences, Protein Translocations and Binding Partners

O’Day

2019

Cells

View full text Add to dashboard Cite

The nucleoli of Dictyostelium discoideum have a comparatively unique, non-canonical, localization adjacent to the inner nuclear membrane. The verified nucleolar proteins of this eukaryotic microbe are detailed while other potential proteins are introduced. Heat shock protein 32 (Hsp32), eukaryotic translation initiation factor 6 (eIF6), and tumour necrosis factor receptor-associated protein 1 (TRAP1) are essential for cell survival. NumA1, a breast cancer type 1 susceptibility protein-C Terminus domain-containing protein linked to cell cycle, functions in the regulation of nuclear number. The cell cycle checkpoint kinase 2 homologue forkhead-associated kinase A (FhkA) and BRG1-associated factor 60a homologue Snf12 are also discussed. While nucleoli appear homogeneous ultrastructurally, evidence for nucleolar subcompartments exists. Nucleolar localization sequences (NoLS) have been defined that target proteins to either the general nucleolar area or to a specific intranucleolar domain. Protein translocations during mitosis are protein-specific and support the multiple functions of the Dictyostelium nucleolus. To enrich the picture, binding partners of NumA1, the most well-characterized nucleolar protein, are examined: nucleolar Ca2+-binding protein 4a (CBP4a), nuclear puromycin-sensitive aminopeptidase A (PsaA) and Snf12. The role of Dictyostelium as a model for understanding the contribution of nucleolar proteins to various diseases and cellular stress is discussed throughout the review.

show abstract

The nucleus of axenically grownDictyostelium discoideum: Studies on its division cycle, isolation and conformation

Cited by 10 publications

References 25 publications

Identification of a UDP-GlcNAc:Skp1-Hydroxyproline GlcNAc-transferase in the Cytoplasm of Dictyostelium

Identification of a UDP-GlcNAc:Skp1-Hydroxyproline GlcNAc-transferase in the Cytoplasm of Dictyostelium

Dictyostelium discoideum Hsp32 is a resident nucleolar heat-shock protein

Proteins of the Nucleolus of Dictyostelium discoideum: Nucleolar Compartmentalization, Targeting Sequences, Protein Translocations and Binding Partners

Contact Info

Product

Resources

About