cDNA cloning of a novel human gene NAKAP95, neighbor of A-kinase anchoring protein 95 (AKAP95) on chromosome 19p13.11–p13.12 region

Seki, Naohiko; Ueki, Nobuhide; Yano, Kazuhiro; Masuho, Yasuhiko; Muramatsu, Masaaki

doi:10.1007/s100380070040

Cited by 12 publications

(9 citation statements)

References 29 publications

(17 reference statements)

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“…Given that, within the nucleolus, ribosomal DNA genes occur in multiple copies (400) per cell and are especially rich in GC-nucleotides within their promoters [46], our results favour the hypothesis that preference for binding to GCrich DNA could be one mechanism that leads to nucleolar accumulation of AKAP95. These results also fit with the consensus for C 2 H 2 zinc-fingers as a DNA-binding protein family [34] that typically displays GC-rich binding preferences [35]. Furthermore, ChIP data reveal that AKAP95 is enriched at rRNA promoters; this finding is entirely consistent with our immunofluorescence analysis showing nucleolar accumulation of AKAP95 at UBFpositive loci in the nucleolus.…”

Section: Discussionsupporting

confidence: 88%

“…AKAP95 preferentially binds GC-rich DNA in vitro and associates to GC-rich ribosomal RNA promoters in vivo Members of the DNA-binding C 2 H 2 zinc-finger protein family [34], typically display GC-rich binding preferences [35]. Because AKAP95 is a C 2 H 2 -type zinc-finger protein, we first investigated whether AKAP95 was able to bind DNA in vitro.…”

Section: Transcription-dependent Targeting Of Akap95 To the Nucleolusmentioning

confidence: 99%

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A‐kinase anchoring protein AKAP95 is a novel regulator of ribosomal RNA synthesis

et al. 2016

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The RNA polymerase I transcription apparatus acquires and integrates the combined information from multiple cellular signalling cascades to regulate ribosome production essential for cell growth and proliferation. In the present study, we show that a subpopulation of A-kinase anchoring protein 95 (AKAP95) targets the nucleolus during interphase and is involved in regulating rRNA production. We show that AKAP95 co-localizes with the nucleolar upstream binding factor, an essential rRNA transcription factor. Similar to other members of the C 2 H 2 -zinc finger family, we show, using systematic selection and evolution of ligands by exponential enrichment and in vitro binding analysis, that AKAP95 has a preference for GC-rich DNA in vitro, whereas fluorescence recovery after photobleaching analysis reveals AKAP95 to be a highly mobile protein that exhibits RNA polymerase I and II dependent nucleolar trafficking. In line with its GC-binding features, chromatin immunoprecipitation analysis revealed AKAP95 to be associated with ribosomal chromatin in vivo. Manipulation of AKAP95-expression in U2OS cells revealed a reciprocal relationship between the expression of AKAP95 and 47S rRNA. Taken together, our data indicate that AKAP95 is a novel nucleolus-associated protein with a regulatory role on rRNA production.

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Section: Discussionsupporting

confidence: 88%

Section: Transcription-dependent Targeting Of Akap95 To the Nucleolusmentioning

confidence: 99%

A‐kinase anchoring protein AKAP95 is a novel regulator of ribosomal RNA synthesis

et al. 2016

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“…ZF, zinc finger. of AKAP95, NAKAP95 (25). Sequence similarities, including a distinct pattern of aromatic residues, were found with AKAP95 from amino acids 127 to 152, and we propose that this region constitutes the NMTS of this protein family.…”

Section: Fig 2 Identification Of a Nuclear Import Signal For Akap95mentioning

confidence: 61%

A-kinase-anchoring Protein AKAP95 Is Targeted to the Nuclear Matrix and Associates with p68 RNA Helicase

Akileswaran

Taraska

Sayer

et al. 2001

Journal of Biological Chemistry

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The cell nucleus is structurally and functionally organized by the nuclear matrix. We have examined whether the nuclear cAMP-dependent protein kinase-anchoring protein AKAP95 contains specific signals for targeting to the subnuclear compartment and for interaction with other proteins. AKAP95 was expressed in mammalian cells and found to localize exclusively to the nuclear matrix. Mutational analysis was used to identify determinants for nuclear localization and nuclear matrix targeting of AKAP95. These sites were found to be distinct from previously identified DNA and protein kinase A binding domains. The nuclear matrix-targeting site is unique but conserved among members of the AKAP95 family. Direct binding of AKAP95 to isolated nuclear matrix was demonstrated in situ and found to be dependent on the nuclear matrix-targeting site. Moreover, Far Western blot analysis identified at least three AKAP95-binding proteins in nuclear matrix isolated from rat brain. Yeast two-hybrid cloning identified one binding partner as p68 RNA helicase. The helicase and AKAP95 co-localized in the nuclear matrix of mammalian cells, associated in vitro, and were precipitated as a complex from solubilized cell extracts. The results define novel protein-protein interactions among nuclear matrix proteins and suggest a potential role of AKAP95 as a scaffold for coordinating assembly of hormonally responsive transcription complexes.The nuclear matrix was first described in 1974 by Berezney and Coffey (1, 2) as the insoluble, proteinaceous structural component of the nucleus that remains after digestion and extraction of chromatin and associated nuclear factors. Since then it has become increasingly apparent that the matrix plays an important functional role, coordinating many nuclear activities including DNA replication, RNA processing, and transcription (3-4). Integration of these events is thought to occur within a high level of chromatin structure where the nuclear matrix organizes DNA into functional loops by binding specific sequences or "matrix attachment regions." Not surprisingly, these regions are often found flanking actively transcribed genes and may serve as localization sites for assembly of factors regulating gene expression (5-7).Structurally, the intact nuclear matrix includes nuclear lamina, residual nucleoli, and ribonucleoprotein particles bound via a fibrous network of proteins, RNA, and polysaccharides. Although lamins and heterogeneous nuclear RNA proteins are the predominant protein constituents, the core structure contains more than 100 distinct nuclear matrix proteins, many of which bind DNA (8 -10). To date, relatively few nuclear matrix proteins have been characterized in detail, and little is known about how they are incorporated into the matrix. Complicating the analysis is the fact that the nuclear matrix is a dynamic structure, and its protein composition is affected by environmental stimuli and growth conditions. Indeed, several nuclear matrix proteins have been identified as useful markers for specific typ...

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“…Chromosomal assignment of the human SOX18 gene was done by polymerase chain reaction (PCR) analysis of a human/rodent somatic cell hybrid panel (National Institute of General Medicine Service, Coriell Cell Repositories, Camden, NJ, USA) and a radiation hybrid panel (Genebridge 4; Research Genetics, Huntsville, AL, USA), as described previously Seki et al 2000). The human SOX18-specific PCR primers (5Ј-GCC AGC AGC GCG GTC TAT TAC-3Ј, nucleotides 1259 to 1279; 5Ј-AGG AAG CGC TGC AGG GAC CCG-3Ј, nucleotides 1316 to 1336), gave rise to an amplified 78-bp product by genomic PCR.…”

Section: Chromosome Mapping Of Human Sox18 Genementioning

confidence: 99%

cDNA cloning, tissue expression, and chromosome mapping of human homolog of SOX18

Abe¹,

Seki²,

Yoshikawa³

et al. 2000

J Hum Genet

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The SRY (sex-determining region Y) gene encodes a transcription factor characterized by a DNAbinding motif termed the HMG (high mobility group) domain. The SOX (Sry-box) genes comprise a large family related by homology to the HMG-box region. We isolated a cDNA clone with an open reading frame encoding a putative protein of 384 amino acids, which shared 83% identity to the mouse Sox18 protein. Northern blot analysis revealed that a 1.9-kb band of human SOX18 messenger RNAs was predominantly expressed in heart, although weak signals were seen in brain, liver, testis, and leukocyte. By polymerase chain reaction (PCR)-based analyses with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid panel, the gene was mapped to the chromosome 20q13.33 region.

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cDNA cloning of a novel human gene NAKAP95, neighbor of A-kinase anchoring protein 95 (AKAP95) on chromosome 19p13.11–p13.12 region

Cited by 12 publications

References 29 publications

A‐kinase anchoring protein AKAP95 is a novel regulator of ribosomal RNA synthesis

A‐kinase anchoring protein AKAP95 is a novel regulator of ribosomal RNA synthesis

A-kinase-anchoring Protein AKAP95 Is Targeted to the Nuclear Matrix and Associates with p68 RNA Helicase

cDNA cloning, tissue expression, and chromosome mapping of human homolog of SOX18

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