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Medizin - Open Access LMU - Teil 11/22 Podcast
Cloning, expression, and chromosomal localization of the 140-kilodalton subunit of replication factor C from mice and humans

Cloning, expression, and chromosomal localization of the 140-kilodalton subunit of replication factor C from mice and humans

Medizin - Open Access LMU - Teil 11/22 vor 32 Jahren

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We have isolated a full-length mouse cDNA encoding a lysine-rich
protein of 1,131 amino acids with a calculated molecular mass of
126 kDa. The protein binds in a sequence-unspecific manner to DNA,
is localized exclusively in the nucleus, and contains a putative
ATP binding site and a stretch of 80 amino acids with homology to
the carboxy terminus of prokaryotic DNA ligases. On the basis of
the following facts, we conclude that the isolated cDNA encodes the
140-kDa subunit of mouse replication factor C (mRFC140). (i) The
sequence around the ATP binding site shows significant homology to
three small subunits of human replication factor C. (ii) Polyclonal
antibodies raised against the protein encoded by this cDNA
cross-react with the 140-kDa subunit of purified human replication
factor C (hRFC140) and recognize in mouse cell extracts an
authentic protein with an apparent molecular mass of 130 kDa. (iii)
Sequence comparison with a human cDNA isolated by using tryptic
peptide sequence information from purified hRFC140 revealed 83%
identity of the encoded proteins. The mRFC140 gene is ubiquitously
expressed, and two mRNAs approximately 5.0 and 4.5 kb long have
been detected. The gene was mapped by in situ hybridization to
mouse chromosome 5, and its human homolog was mapped to chromosome
4 (p13-p14).