Human DNase I Contains Mannose 6-Phosphate and Binds the Cation-Independent Mannose 6-Phosphate Receptor

Cacia, Jerry; Quan, Cynthia; Pai, Roger; Frenz, John

doi:10.1021/bi981465t

Cited by 16 publications

(7 citation statements)

References 71 publications

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“…Another probe used to detect nuclear monomeric or G-actin is labeled DNase I, a serum deoxyribonuclease [49]. DNase I binds to ATP-G-actin with high affinity, although it also binds F-actin [83,84], DNA [85,86], and other proteins [87]. In conclusion, neither reagent is highly efficient for the study of nuclear actin.…”

Section: Actin Staining In Fixed Cellsmentioning

confidence: 99%

Nuclear Actin and Actin-Binding Proteins in DNA Repair

Hurst

Shimada

Gasser

2019

Trends in Cell Biology

115

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Nuclear actin has been implicated in a variety of DNA-related processes including chromatin remodeling, transcription, replication, and DNA repair. However, the mechanistic understanding of actin in these processes has been limited, largely due to a lack of research tools that address the roles of nuclear actin specifically, that is, distinct from its cytoplasmic functions. Recent findings support a model for homology-directed DNA double-strand break (DSB) repair in which a complex of ARP2 and ARP3 (actin-binding proteins 2 and 3) binds at the break and works with actin to promote DSB clustering and homology-directed repair. Further, it has been reported that relocalization of heterochromatic DSBs to the nuclear periphery in Drosophila is ARP2/3 dependent and actin-myosin driven. Here we provide an overview of the role of nuclear actin and actin-binding proteins in DNA repair, critically evaluating the experimental tools used and potential indirect effects. Nuclear Actin: Globular and Filamentous Forms Highlights Methods used to study nuclear actin have major caveats that can lead to biased data interpretation. Globular actin in nuclear remodeling complexes and histone acetyltransferases is often neglected due to a focus on filamentous nuclear actin. Recent evidence implicates nuclear actin and actin-binding proteins in resectiondependent DNA repair. The mechanism is unclear. The field needs further development of tools, the use of technical controls, and a systematic evaluation of alternative interpretations to reveal the function of nuclear actin.

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Section: Actin Staining In Fixed Cellsmentioning

confidence: 99%

Nuclear Actin and Actin-Binding Proteins in DNA Repair

Hurst

Shimada

Gasser

2019

Trends in Cell Biology

115

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“…In addition to the known acid hydrolases, bovine pancreatic DNase I was tested as a substrate because it has been shown to be phosphorylated by GlcNAc-1-phosphotransferase in vivo (34). It interacted with both forms of the transferase with good affinity (K m,app of 30 M for ␣ 2 ␤ 2 ␥ 2 and 60 M for ␣ 2 ␤ 2 ) and similar k cat values, although these values were considerably lower than those obtained with the acid hydrolases ( Table 1).…”

Section: Acceptormentioning

confidence: 99%

Functions of the α, β, and γ Subunits of UDP-GlcNAc:Lysosomal Enzyme N-Acetylglucosamine-1-phosphotransferase

Qian

Lee

et al. 2010

Journal of Biological Chemistry

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UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase is an ␣ 2 ␤ 2 ␥ 2 hexamer that mediates the first step in the synthesis of the mannose 6-phosphate recognition marker on lysosomal acid hydrolases. Using a multifaceted approach, including analysis of acid hydrolase phosphorylation in mice and fibroblasts lacking the ␥ subunit along with kinetic studies of recombinant ␣ 2 ␤ 2 ␥ 2 and ␣ 2 ␤ 2 forms of the transferase, we have explored the function of the ␣/␤ and ␥ subunits. The findings demonstrate that the ␣/␤ subunits recognize the protein determinant of acid hydrolases in addition to mediating the catalytic function of the transferase. In mouse brain, the ␣/␤ subunits phosphorylate about one-third of the acid hydrolases at close to wild-type levels but require the ␥ subunit for optimal phosphorylation of the rest of the acid hydrolases. In addition to enhancing the activity of the ␣/␤ subunits toward a subset of the acid hydrolases, the ␥ subunit facilitates the addition of the second GlcNAc-P to high mannose oligosaccharides of these substrates. We postulate that the mannose 6-phosphate receptor homology domain of the ␥ subunit binds and presents the high mannose glycans of the acceptor to the ␣/␤ catalytic site in a favorable manner.In higher eukaryotes, newly synthesized acid hydrolases acquire mannose 6-phosphate (Man-6-P) 3 residues on their N-linked glycans as they traverse the Golgi (1). These residues serve as high affinity ligands for binding to Man-6-P receptors in the trans-Golgi network. The hydrolase-receptor complexes are then packaged into transport carriers for delivery to endosomes and lysosomes. The Man-6-P recognition marker is synthesized in two steps. First, UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-1-phosphotransferase) binds to a conformation-dependent protein determinant on the acid hydrolase and transfers GlcNAc-1-P from UDP-GlcNAc to one or two of the mannose residues of the N-linked high mannose oligosaccharide. Second, N-acetylglucosamine 1-phosphodiester ␣-N-acetyglucosaminidase ("uncovering enzyme") excises the N-acetylglucosamine to generate the Man-6-P monoester.GlcNAc-1-phosphotransferase is a heterohexamer composed of three subunits (␣ 2 ␤ 2 ␥ 2 ) (2). The ␣ and ␤ subunits are encoded by a single gene GNPTAB, and the ␥ subunit is encoded by a separate gene GNPTG (3-5). Although it has been established that the ␣/␤ subunits contain the catalytic activity of the enzyme, the possible participation of these subunits in the recognition of the common protein determinant of the acid hydrolases has not been explored. Furthermore, the role(s) of the ␥ subunit is poorly understood. The initial insight into the function of the subunits came from studies of patients with the autosomal recessive lysosomal storage disorders termed mucolipidosis II (I-cell disease) and mucolipidosis III (pseudo-Hurler polydystrophy), the latter being the less severe of the two (6). These disorders arise from mutations in the genes encoding GlcNAc-1-phospho...

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“…To confirm the hypothesis that DNase I can enter into living cells upon engagement of a cell surface molecule able to internalize it, we first sought to define the membrane receptor for DNase I. DNase I is a glycoprotein, contains mannose 6‐phosphate residues, and has been reported to bind the cation‐independent mannose 6‐phosphate/insulin‐like growth factor receptor (CI‐MPR) immobilized to a column 10. CI‐MPR is a type I glycoprotein with high affinity for proteins that contain phosphomannosyl residues 11.…”

Section: Resultsmentioning

confidence: 99%

DNase I behaves as a transcription factor which modulates Fas expression in human cells

et al. 2003

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DNase I is the major nuclease present in biological fluids and is ubiquitously expressed in mammalian tissues. It is responsible for the removal of DNA from nuclear antigens, and consistently with this function, DNase I-deficient mice show features of autoimmunity. The enzyme seems also to be involved in apoptosis (programmed cell death). We demonstrate that DNase I is internalized by human cells upon binding mannose 6-phosphate receptor and gains access into the cells. Following internalization of the enzyme, the cells show an increased surface expression of Fas molecule, a key regulator of apoptosis. Here we show that DNase I up-regulates fas transcription upon interaction with the fas gene promoter. Moreover, overexpression of the DNase I gene in human cells results in a similar modulation of the fas gene expression. Our data provide the first evidence that the endonuclease DNase I behaves as a transcription factor which selectively regulates cell surface Fas expression in human cells and point towards a fundamental role of DNase I in the regulation of the apoptotic machinery.

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Human DNase I Contains Mannose 6-Phosphate and Binds the Cation-Independent Mannose 6-Phosphate Receptor

Cited by 16 publications

References 71 publications

Nuclear Actin and Actin-Binding Proteins in DNA Repair

Nuclear Actin and Actin-Binding Proteins in DNA Repair

Functions of the α, β, and γ Subunits of UDP-GlcNAc:Lysosomal Enzyme N-Acetylglucosamine-1-phosphotransferase

DNase I behaves as a transcription factor which modulates Fas expression in human cells

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