Cytotoxic ribonucleases: molecular weapons and their targets

Makarov, Alexander А.; Ilinskaya, О. N.

doi:10.1016/s0014-5793(03)00225-4

Cited by 163 publications

(148 citation statements)

References 57 publications

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“…7, 8). It should be noted that some RNAses with cell-penetrating activity, such as Onconase and bovine seminal RNase, also induced apoptotic cell death by damaging cytosolic RNAs [34,40]. The present work shows that 3D8 scFv translocates across the plasma membrane by caveolae/lipid-raftmediated endocytosis upon interacting with anionic proteoglycans on the cell surface, accumulates in the cytosol, and then induces apoptotic cell death by hydrolyzing cellular RNAs.…”

Section: Discussionmentioning

confidence: 51%

A nucleic acid-hydrolyzing antibody penetrates into cells via caveolae-mediated endocytosis, localizes in the cytosol and exhibits cytotoxicity

Jang

Jeong

Jun

et al. 2009

Cell. Mol. Life Sci.

122

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Abstract. Many natural anti-DNA antibodies (Abs) have the ability to translocate across the plasma membrane and localize in the nucleus of mammalian cells, frequently leading to cytotoxicity to cells. Herein, we report detailed intracellular trafficking routes and cytotoxicity in HeLa cells for a single chain variable fragment (scFv) Ab, 3D8, which is an anti-DNA Ab capable of hydrolyzing both DNA and RNA. The intracellular penetration of 3D8 scFv occurred by caveolae/lipid raft endocytosis. The time-course chasing experiments revealed that 3D8 scFv escaped directly from the caveosome into the cytosol and remained in the cytosol without further trafficking into endosomes, lysosomes, endoplasmic reticulum, Golgi, or nucleus. The cytosolically localized 3D8 scFv maintained its nuclease activity to hydrolyze cellular RNAs, mainly mRNAs, eventually triggering apoptotic cell death. Our results demonstrate that 3D8 scFv has a unique intracellular trafficking route of localizing in the cytosol, thereby exhibiting cytotoxicity due to its nuclease activity.

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Section: Discussionmentioning

confidence: 51%

A nucleic acid-hydrolyzing antibody penetrates into cells via caveolae-mediated endocytosis, localizes in the cytosol and exhibits cytotoxicity

Jang

Jeong

Jun

et al. 2009

Cell. Mol. Life Sci.

122

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“…Binase is a purine-specific RNase of T1 family and a typical representative of the microbial RNases. 5,15,17 Earlier the cytotoxic activity of a number of microbial RNases, including binase, was shown in vitro on different types of tumor cells, 1,5,[9][10][11][28][29][30] but no studies have been performed in vivo.…”

Section: Discussionmentioning

confidence: 99%

Ribonuclease binase inhibits primary tumor growth and metastases via apoptosis induction in tumor cells

et al. 2013

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“…Paradigmatic examples are bovine seminal RNase (BSRNase) 1,2 and RNase from oocytes of Rana pipiens [3][4][5] (commercial trademark of Alfacell, USA, onconase (ONC)), both belonging to the pancreaticlike RNase family, whose prototype is the bovine pancreatic enzyme (RNase A). 6,7 This unusual biological activity is critically dependent on the ribonucleolytic activity 8 and on the ability of these molecules to reach the cellular cytosol and to degrade RNA by evading the action of the ribonuclease inhibitor (RI). 9,10 It is currently accepted that RI, which binds most RNase A family members with femtomolar affinities, plays an important role in the protection of host cells from endogenous RNases.…”

Section: Introductionmentioning

confidence: 99%

Structural features for the mechanism of antitumor action of a dimeric human pancreatic ribonuclease variant

et al. 2008

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A specialized class of RNases shows a high cytotoxicity toward tumor cell lines, which is critically dependent on their ability to reach the cytosol and to evade the action of the ribonuclease inhibitor (RI). The cytotoxicity and antitumor activity of bovine seminal ribonuclease (BSRNase), which exists in the native state as an equilibrium mixture of a swapped and an unswapped dimer, are peculiar properties of the swapped form. A dimeric variant (HHP2-RNase) of human pancreatic RNase, in which the enzyme has been engineered to reproduce the sequence of BSRNase helix-II (Gln28fiLeu, Arg31fiCys, Arg32fiCys, and Asn34fiLys) and to eliminate a negative charge on the surface (Glu111fiGly), is also extremely cytotoxic. Surprisingly, this activity is associated also to the unswapped form of the protein. The crystal structure reveals that on this molecule the hinge regions, which are highly disordered in the unswapped form of BSRNase, adopt a very well-defined conformation in both subunits. The results suggest that the two hinge peptides and the two Leu28 side chains may provide an anchorage to a transient noncovalent dimer, which maintains Cys31 and Cys32 of the two subunits in proximity, thus stabilizing a quaternary structure, similar to that found for the noncovalent swapped dimer of BSRNase, that allows the molecule to escape RI and/or to enhance the formation of the interchain disulfides.Keywords: crystal structure; antitumor agents; ribonuclease; 3D-domain swapping; dimers Abbreviations: BSRNase, bovine seminal ribonuclease; des(1-7)HP-RNase, human pancreatic ribonuclease in which the first seven residues have been deleted; HHP-RNase, covalent dimeric variant of human pancreatic ribonuclease in which four residues at the helix-II region are mutated according to BSRNase sequence (Gln28!Leu, Arg31!Cys, Arg32!Cys, and Asn34!Lys); HHP2-RNase, HHP-RNase with an additional mutation (Glu111!Gly); HP-RI, structure of human pancreatic ribonuclease complexed with ribonuclease inhibitor; HP-RNase, human pancreatic ribonuclease; MxM-BSRNase, dimeric form of BSRNase in which the chains swap their N-terminal tails; M¼M-BSRNase, unswapped dimer of BSRNase; M¼M-HHP2, unswapped dimer of HHP2-RNase; NCD-BSRNase, noncovalent swapped form of BSRNase; ONC, Onconase; PDB, Protein Data Bank; PM7, human pancreatic ribonuclease in which residues 1-21 are replaced by the corresponding residues of BSRNase; PM8, N-terminal swapped dimer of a variant of human pancreatic ribonuclease; RI, ribonuclease inhibitor; RMSD, root-mean-square deviation.Grant sponsor: Ministero dell'Istruzione, dell'Università e della Ricerca; Grant number: FIRB RBNE03B8KK and PRIN2007.

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Cytotoxic ribonucleases: molecular weapons and their targets

Cited by 163 publications

References 57 publications

A nucleic acid-hydrolyzing antibody penetrates into cells via caveolae-mediated endocytosis, localizes in the cytosol and exhibits cytotoxicity

A nucleic acid-hydrolyzing antibody penetrates into cells via caveolae-mediated endocytosis, localizes in the cytosol and exhibits cytotoxicity

Ribonuclease binase inhibits primary tumor growth and metastases via apoptosis induction in tumor cells

Structural features for the mechanism of antitumor action of a dimeric human pancreatic ribonuclease variant

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