Three genes (sspH, sspL, and tlp) encoding new, minor small, acid-soluble proteins (SASP) unique to spores of Bacillus subtilis are expressed only in the forespore compartment during sporulation of this organism. The sspH and sspL genes are monocistronic, whereas tip is the second gene in an operon with a second small orf, which we have termed sspN. The sspN and sspL genes are recognized primarily by the forespore-specific sigma factor for RNA polymerase, sigma(G); the sspN-tlp operon is recognized equally well by sigma(G) and the other forespore-specific sigma factor, sigma(F). Sequences centered 10 and 35 nt upstream of the 5'-ends of sspH, sspL, and sspN mRNAs all show homology to -10 and -35 sequences recognized by sigma(F) and sigma(G), which are generally quite similar. Mutations disrupting the sspH, sspL, sspH-tlp, or tlp loci cause a loss of the appropriate SASP from spores, but have no discernible effect on sporulation, spore properties, or spore germination. (C) 1999 Elsevier Science B.V. All rights reserved.