The Immunogen : Nsr1p is localized in the nucleolus and binds to telomeric single-stranded oligonucleotide repeat sequence of the type and binds to the single-stranded telomeric repeat sequence (TG1-3)n. It has a role in ribosomal RNA maturation and possibly in transport of proteins into the nucleus.

Diagram of Domain Structure: Generated from sequence of yeast Nsr1p with the SMART program from EMBL in Heidleberg. The first block from the left is a glutamic acid rich region predicted by SMART to encode a region of a-helical coiled coil, though this is likely to be an artifact of the algorithm used. The blocks marked "RRM" indicate two RRM RNA binding domains, each about 70 amino acids long. RRM stands for "RNA recognition motif" and the domain, found in many eukaryotic proteins which bind single stranded RNAs, is also known as the putative RNA-binding region RNP-1 signature. The final block is a region of low sequence complexity, in this case rich in arginine and glycine residues Scale is number of amino acids;

Antibody Characteristics: MCA-31C4 was isolated during a screen for monoclonal antibodies reactive with yeast nucleolar proteins. We determined the specificity by screening a yeast genomic library expressed as b-galactosidase fusion proteins in lgt11. Two individually isolated, different positive clones were found. Sequencing revealed that in both clones b-galactosidase was fused at the EcoRI site to the amino acid sequence beginning with …EFEH, corresponding to amino acids 190 and onwards in Nsr1p. This peptide is ~20 amino acids into the first RRM domain. Thus, MCA-31C4 reacts with an epitope in the C-terminal 55% of Nsr1p. The antibody is provided as sterile-filtered cell culture fluid from an Integra CL-350 biochamber plus sodium azide. The IgG isotype and exact concentration are not known.

Suggestions for use: For western blots of yeast protein samples (cell and nuclear lysates), try MCA-31C4 diluted 1/10,000, followed by chemiluminescent detection (ECL). For non-ECL western detection methods, try MCA-31C4 diluted 1/1,000 to 1/5,000. Press here for image of blot. For immunofluorescence on yeast cells, try MCA-31C4 diluted 1/500 to 1/5,000. Antibody preparation contains 10mM sodium azide preservative (Press here for Material Safety Data Sheet). Avoid repeated freezing and thawing, store at 4°C or -20°C.

UniProt/Swiss-Prot Link: press here

References:

Lin JJ and Zakian VA. Isolation and characterization of two Saccharomyces cerevisiae genes that encode proteins that bind to (TG1-3)n single strand telomeric DNA in vitro. Nucleic Acids Res 22:4906-13 1994.

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