Subbulakshmi (Kunjamma to her family) was born on September 16, 1916 in Madurai, Madras Presidency, India to veena player Shanmukavadiver Ammal and Subramania Iyer. Her grandmother Akkammal was a Violinist.

Subbulakshmi gave her first public performance, at the age of eleven, in the year 1927, in the 100 pillar hall inside the Rockfort Temple, Tiruchirappalli; with Mysore Chowdiah on the violin and Dakshinamurthy Pillai on the mridangam. This was organised by the Tiruchirappalli based Indian National Congress leader F. G. Natesa Iyer.

Subbulakshmi gave her first performance at the prestigious Madras Music Academy in 1929, when she was 13 years old . The performance consisted of singing bhajans (Hindu hymns). The academy was known for its discriminating selection process, and they broke tradition by inviting a young girl as a key performer. Her performance was described as spellbinding and earned her many admirers and the moniker of musical genius from critics. Soon after her debut performances, Subbulakshmi became one of the leading Carnatic vocalists.

In 1936 Subbulakshmi moved to Madras (now Chennai). She also made her film debut in Sevasadan in 1938. Her debut to the world of cinema was again opposite F. G. Natesa Iyer: who had introduced her as a child Prodigy to the world of Carnatic music a decade earlier, in Tiruchirappalli.

M.S. also acted in a few Tamil films in her youth. Her first movie, Sevasadanam, was released on 2 May 1938. F.G. Natesa Iyer was the lead actor, opposite Subbulakshmi, in this film, directed by K. Subramanyam. It was a critical and commercial success. Ananda Vikatan favourably reviewed the film on 8 May 1938:

MS Subbulakshmi also played the male role of Narada in "Savitri" (1941) to raise money for launching Kalki, her husband's nationalist Tamil weekly. Her title role of the Rajasthani saint-poetess Meera in the eponymous 1945 film gave her national prominence. This movie was re-made in Hindi in 1947.

were significant landmarks in her career. In 1969 she was accompanied by Indian Railways Advisor SN Venkata Rao to Rameshwaram, where she famously sang several songs in front of each idol in the Rameshwaram temple.

After the death of her husband Kalki Sadasivam in 1997, she stopped all her public performances.

A commemorative postage stamp on her was issued on 18-December-2005. United Nations decided to issue stamp to mark birth centenary M.S. Subbulakshmi, She was bestowed with enormous prize moneys with these awards, most of which she donated to charity. She has given more than 200 charity concerts and raised well over Rs. 10,000,000. She was awarded honorary degrees from several Universities. She was an ardent devotee of Kanchi Mahaswamigal and she rendered his composition Maithreem Bhajatha (O World! Cultivate peace) in her concert at the UN in 1966. She made a 20-minute recording of Venkatesa Suprabhatam for HMV, the royalty from which goes to the Veda Patasala run by the Tirupati Tirumala Devasthanam. She donated many of the royalties on several best sold records to many charity organisations.

She was honoured as a resident Artist [Asthana Vidhwan] of Tirumala Tirupati Devasthanams. Tirupati Urban Development Authority (TUDA) has installed a bronze statue of M.S. Subbulakshmi at the Poornakumbham circle in the temple town. It was unveiled by Andhra Pradesh Chief Minister Y. S. Rajasekhara Reddy on 28 May 2006.