TABLE 2

Overexpressed cDNA clones disrupt telomeric silencing

Gene (ORF size)ClonesInsert size (kb)Predicted expressed region (aa)Comments
SIR4 (1358 aa)pTRP2, pTRP31.271009–1358
pTRP40.77 b
pTRP8, pTRP320.791242–1358 c
pTRP10,a pTRP42, pTRP46, pTRP521.131071–1358
pTRP161.471009-1358
pTRP580.85 d
ASF1 (279 aa)pTRP90.921–279
pTRP19, pTRP410.991–279
pTRP22, pTRP36, pTRP630.981–279
pTRP17, pTRP31, pTRP48, pTRP53a1.041–279
pTRP301.001–279 e
pTRP621.021–279
pTRP570.951–279
IFH1 (1085 aa)pTRP1, pTRP340.801–213
pTRP70.921–218
pTRP130.911–212
pTRP20a0.921–216
pTRP270.921–212
DNA2 (1522 aa)pTRP351.071–338
pTRP56a1.091–337
RPL32 (105 aa)pTRP240.461–105
pTRP54a0.901–105 f
TLC1 (1.3-kb functional RNA)pTRP6a1.25 g
pTRP611.25
pTRP14, pTRP471.21
pTRP33, pTRP391.22
pTRP551.21
pTRP591.21
pTRP601.00
DOT1 (582 aa)pTRP28a1.881–582
DOT4 (792 aa)ipTRP50a2.4316–784 (with respect to GenBank Sequence) h
DOT5 (215 aa)pTRP49a0.811–215
DOT6 (670 aa)pTRP290.891–286
pTRP23a2.1237–670
  • a The plasmids used as representative clones for the overexpression studies.

  • b Antisense. RNA sequence is complementary to part of the SIR4 mRNA: the last 593 nt of the SIR4 ORF and 172 nt of the 3’ untranslated region.

  • c The first ORF in the cDNA is a 35-amino-acid-encoding sequence in a different reading frame.

  • d Antisense. RNA sequence is complementary to part of the SIR4 mRNA: the last 658 nt of the SIR4 ORF and 189 nt of the 3’ untranslated region.

  • e Fused to 75-nt fragment of SUM1 antisense RNA.

  • f 3’ end of RPL32 RNA is fused to RPS24 RNA (starting at nucleotide 30 of the RPS24 ORF).

  • g All the TLC1 clones have been described previously (Singer and Gottschling 1994).

  • h pTRP50 has four point mutations and encodes a different C-terminal tail compared to the DOT4 sequence contained in GenBank. After amino acid 784, the GenBank protein sequence terminates with the peptide NKKRKFTK. The protein encoded by pTRP50 terminates with the sequence IKKGSSPNEKTRYSWIFLFS.

  • i GenBank sequence.