A growth-responsive gene (16C8) in normal mouse fibroblasts homologous to a human collagenase inhibitor with erythroid-potentiating activity: evidence for inducible and constitutive transcripts

Abstract

We present the DNA sequence of an essentially full-length cDNA clone of 16C8, a growth factor-inducible gene isolated from a mouse embryo fibroblast cDNA library. The 0.9-kb mRNA encodes an Mr 22,500 protein that has substantial homology to a human protein with the reported abilities (1) to potentiate erythroid differentiation and (2) to inhibit collagenases and other tissue metalloproteinases. The N-terminus of the predicted protein has a hydrophobic nature characteristic of secreted proteins, and two potential sites for Nlinked glycosylation are present. The cytoplasmic concentration of 16C8 mRNA is maximal in mid Gl at about 6 h after serum stimulation of quiescent fibroblasts. Northern blot analysis showed a progressive reduction in the size of the induced 16C8 transcripts with increasing time after serum stimulation. This was shown to be due to the reduction in length of the poly(A) tails. S1 analysis of the 5' portion of the mRNA revealed the presence of three different species of transcript, only one of which was inducible.

Murine tissue inhibitor of metalloproteinases—4 (Timp—4): cDNA isolation and expression in adult mouse tissues1

“…In contrast to the patterns seen with Timp-\, -2 and -3, expression of Timp-4 mRNA was restricted to brain, heart, ovaries, skeletal muscle and skin. In other studies, we failed to detect expression of Timp-4 by Northern blot analysis in normal or Ha-rastransformed C3H10T1/2 mouse fibroblasts, though the other three Timp genes showed characteristic cell line-specific and stimulus-responsive patterns of expression [19,21,22]. …”

“…Following transformation, bacteria were grow on LB agar containing 50 μg/ml ampicillin and colonies were transferred to nylon filters (Hybond N, Amersham). Filters were hybridized with a mixture of [ 32 P]dCTP-labelled Timp-1, Timp-2, and Timp-3 cDNA probes [19,21,22] and washed as previously described [19]. Dried filters were exposed to XAR5 X-ray film (Eastman Kodak).…”

“…Comparison of predicted amino acid sequences of mouse TIMPs. The sequences used for comparison include TIMP-1 [21,28] (these sequences are identical and differ at nine positions from another mouse TIMP-1 sequence in [29]); TIMP-2 [30], chosen in preference to [22] as there are two differences (H and E at positions 12 and 195) in which the chosen mouse sequence shows a better match with human TIMP-2; TIMP-3 [19]. The shaded areas represent 75% identity.…”

Gene encoding a novel murine tissue inhibitor of metalloproteinases (TIMP), TIMP‐3, is expressed in developing mouse epithelia, cartilage, and muscle, and is located on mouse chromosome 10

“…6A,B), pectorals, trapezius, the diaphragm (Fig. 8A,B) and and mouse TIMP-1 (Edwards et al, 1986, Gewert et al , 1987. Gaps introduced in the sequences for maximal alignment are represented by dashes Boxed and shaded areas indicate amino acid residues conserved among all the sequences shown paraspinal muscles.…”

Cell-cycle-specific and serum-dependent expression of gamma-actin mRNA in Swiss mouse 3T3 cells.

“…Edwards et al (9), using approaches similar to those used by others, have characterized five Gl-specific genes in Swiss mouse 3T3 cells. Subsequently, one of these genes was reported to share sequence homology with a gene for human collagenase inhibitor (10).…”